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Objective: To study the effects of different inoculation periods on seedling growth and steroidal saponin content of Paris polyphylla var. yunnanensis, in order to lay a foundation for cultivating and domesticating high-quality Paris polyphylla var. yunnanensis. Methods: The content of steroidal saponins I, II, VI, and VII in the different vaccination periods of Paris polyphylla var. yunnanensis was determined by HPLC. Meanwhile, trypan blue, UV spectrophotometry and so on were adopted to explore colonization rate, colonization intensity, physiological and biochemical indexes, rhizome biomass of the roots of Paris polyphylla var. yunnanensis. Results: Infection rate of AM fungi was higher in different inoculation periods, activities of protective enzymes, photosynthetic pigments and soluble sugars were increased, soluble protein was not changed, content of MDA was decreased, stress resistance of the seedlings of Paris polyphylla var. yunnanensis was improved, and the growth and development were good. Quality of the cultivar one-year seedlings (recovered in August 2015, T7) was relatively low, and cultivar one-year seedlings (harvested in June or July 2015, T5 and T6) and cultivar two years seedlings (recovered in August 2015, T8) were the best. S2, S3 and S6 treatment groups had better effect in different AM fungi mixed groups. Conclusion: Seedling growth and steroidal saponin content of Paris polyphylla var. yunnanensis inoculated by different inoculation periods had certain effects.
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Objective To screen preponderant arbuscular mycorrhizal (AM) fungi species from Paris polyphylla var. yunnanensis seedlings based on mycorrhiza viability, rhizome biomass, and active components. Methods After 28 exogenous species of AM fungi were inoculated in sterilized soil, the effects of the mycorrhiza viability of seedling, the biomass in rhizome, the total amount of polyphyllins in single whole seedling, the amount of polyphyllins in rhizome, and the amount of nucleosides in rhizome of P. polyphylla var. yunnanensis were observed by inoculation test in pot at room temperature. Results All the 28 AM groups formed well mycorrhizal structure, the mycorrhiza viability of P. polyphylla var. yunnanensis seedling was enhanced, but it showed preference. Most of the AM treatment groups could improve the biomass in rhizome, the contents and yields of polyphyllins increased, which implied that the accumulation of polyphyllins and nucleosides in P. polyphylla var. yunnanensis seedling were enhanced after inoculation. AM also changed polyphyllins and nucleosides allocation in different organs of P. polyphylla var. yunnanensis seedling in the secondary metablism, the total amount of four kinds of polyphyllins in rootlet were higher than those in rhizome, but the total amount of nine kinds of nucleosides in rhizome were higher than rootlet. Conclusion The performance of different AM strains in this study is of great reference for further improving the biological quantity and quality of the screening of preponderant AM fungi species from P. polyphylla var. yunnanensis seedlings. Gigaspora gigantean, Scutellospora calospora, Diversispora spurca, Dentiscutata heterogama, Septoglomus deserticola, and Ambispora leptoticha can use as the preponderant AM fungi species for the cultivation of mycorrhizal seedlings of P. polyphylla var. yunnanensis.
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Objective To provide a reference for the introduction and cultivation of Paris polyphylla var. yunnanensis in mid-south region of China, its resistance adaptation ability of high temperature was explored. Methods Strong seedlings of P. polyphylla var. yunnanensis were chosen and treated under different temperature conditions (25 ℃/20 ℃, 33 ℃/28 ℃, 39 ℃/34 ℃) for 7 d, and then investigated the effect on their related physiological indexes. Results The results showed that with the increase of temperature and stress time, the relative water content of the leaves of P. polyphylla var. yunnanensis decreased significantly under the treatment of 33 ℃/28 ℃ and 39 ℃/34 ℃ for 7 d, which was 17.38% and 58.25% lower than the control (25 ℃/20 ℃) respectively. The relative conductivity and the content of malondialdehyde (MDA) were both increased continuously, and it was respectively 3.59 times and 2.55 times higher than that of control under the temperature of 39 ℃/34 ℃. The content of soluble sugar decreased gradually at 39 ℃/34 ℃, which was 33.25% lower than that of the control. The changing of soluble protein content showed a wavy trend, and the fluctuation range of 33 ℃/28 ℃ treatment groups was larger than that of the control group, which increased to 1.32 times of the control after 7 d. The SOD increased firstly and then decreased. The peak value was reached after 4 d at 39 ℃/34 ℃, which was 2.35 times higher than that of the control group. The POD increased gradually in treatment group, which increased to 2.33 times of that in control group at 39 ℃/34 ℃ for 4 d, and the amplitude of the change gradually slowed down. Conclusion In the experimental temperature range, P. polyphylla var. yunnanensis grew well at 25 ℃/20 ℃, which was its optimal growth temperature. Long-term continuous temperature stress treatment at 33 ℃/28 ℃ can cause damage to plant leaves; High temperature at 39 ℃/34 ℃ will seriously damage the physiological and morphological structure of the plant, and even lead to the death of the plant.
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Objective: The genetic diversity of Paris polyphylla var. yunnanensis germplasmic resources in natural populations was analyzed by simple sequence repeat marker for protection and rational utilization. Methods: A total of 115 individuals of P. polyphylla var. yunnanensis collected from five natural populations were analyzed by SSR marker for genetic diversity analysis. Results: Eight pairs of SSR primers were screened. A relatively high level of genetic diversity was revealed: The percentage of polymorphic bands was 100%, the polymorphism information content was 0.745 6. At the population level and species level, the observed number of allele was 8.425 0 and 17.750 0, respectively, the effective number of allele was 4.960 9 and 7.500 7, respectively, the observed heterozygosity was 0.295 5 and 0.294 8, respectively, the expected heterozygosity was 0.654 8 and 0.774 4, respectively, and the Shannon's information index was 1.520 1 and 2.038 6, respectively. The genetic differentiation coefficient was 0.172 8 and the gene flow was 1.196 6. Based on UPGMA cluster analysis, the five populations of P. polyphylla var. yunnanensis were divided into two clades. Conclusion: The level of genetic diversity of P. polyphylla var. yunnanensis was relatively high and there was certain genetic differentiation within and among populations. This study puts forward several suggestions for the protection and development of P. polyphylla var. yunnanensis resources, which will be helpful for protection and sustainable utilization of scientific advice and reference.
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Objective: To explore the impacts of temperature, endogenous hormone, and pollination methods on the seeds germination and emergence of Paris polyphylla var. yunnanensis and polygerm varieties, and establish the technical system to promote the germination of P. polyphylla var. yunnanensis seeds and provide scientific guidance for the cultivation of Paris L., To ease the bottleneck problem of current serious shortage of high quality source of seeding material and commercial herbal products of Paris L., and highest market price. Methods: Using wet sand cultured P. polyphylla var. yunnanensis seeds under different conditions and determined the rate of germination and the mean germination time, the cultivated seeds were planted in the field to investigate the rate of seedling emergence and the average time of emergence. Results: The higher germination rate and emergence rate were obtained at the temperature of 4-18℃; Gibberellin (GA3) treatment could promote the seed germination, and reduce the mean germination time, especially in 100 and 200 mg/L; The germination rate and emergence rate of artificial pollination were significantly higher than natural pollination (P < 0.05); There is no difference between P. polyphylla var. yunnanensis and polygerm varieties seeds. Conclusion: The peeled seeds are treated by 100 mg/L gibberellin, and cultured to sprout by wet sand at the temperature of 4-18℃, then sown in the field. It can shorten the seed emergence period one year and up to more than 80% of emergence rate.
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Objective: To study the rule of breaking seed dormancy and content changes of endogenous hormone of the Paris polyphylla var. Yunnanensis seeds. Methods: The seeds were treated under fluctuating temperature, and the content of endo-hormone, such as GA, IAA, and ABA, was mensurated by HPLC. Results: During the first three months, the stratification under fluctuating temperature could break the seed dormancy. The content of GA rose obviously and the content of ABA reduced. Conclusion: It is suggested that ABA and GA have significant effect on the dormancy and germination of P. polyphylla var. yunnanensis seeds.