| Rationales: Paris polyphylla Smith var.chinensis is a liliaceous perennial of medicinal plant,due to the pharmacological action,to Chinese medicine decoction pieces and traditional Chinese medicine drugs preparations are commonly used in clinical and related industries.Famous historical preparations,such as Yunnan bai yao,Jidesheng snake medicine,Gongxuening capsule,Kangbingdu Granules are commonly used to relate diseases.However,Paris polyphylla Smith var.chinensis needs several years to grow and special requirements for the environment.Paris polyphylla Smith var.chinensis has a long growth cycle and requires high environment requirements for growth.Today,it relies mainly on the supply of wild resources.With over-exploitation over the years,Paris polyphylla Smith var.chinensis resources have gradually dried up,and the conflicts between supply and demand have become increasingly prominent.Seeds are the most suitable method for large-scale and standardized production in the artificial breeding mode of production.However,there are some problems in seed production,such as the uncertainty of seed sources,lack of seed test standards,poor plant lodging resistance,and no systematic evaluation of the effect of different breeding environments on the quality of medicinal materials.Therefore,this study conducted a systematic and extensive research on issues related to Paris polyphylla Smith var.chinensis,in order to solve some problems in the actual production,and provide scientific basis for the standardization of Paris polyphylla Smith var.chinensis.Objective: 1.Combining with proven testing methods and procedures,the methods applied to Paris polyphylla Smith var.chinensis seed quality testing were explored and a scientific,reasonable and effective inspection system was developed for the purpose of evaluating the quality of Paris polyphylla Smith var.chinensis seed production,use,and circulation,so as to improve the lack of previous inspection standards.2.In the actual growth process of Paris polyphylla Smith var.chinensis,will appear the same primordial primordium produces different traits,and there are similar traits in different basal primitives.The problem of Paris polyphylla Smith var.chinensis provenance chaos was solved through the combination of the primordial research,morphological classification and DNA barcoding techniques.3.During the growth of Paris polyphylla Smith var.chinensis,the plants grew too fast and too high,they were prone to lodging,which caused a great influence on the maturity and seed harvest at the later stage of seed breeding.Different plant growth regulators were used to improve the growth of the plant stems,and by reducing the lodging resistance of the plants,the reduction in yield due to poor lodging resistance was reduced.4.By measuring the active ingredients changes of Paris polyphylla Smith var.chinensis in different environments,the effects of different breeding environments on different Paris polyphylla Smith var.chinensis harvesting sites were evaluated systematically,which provided a theoretical basis for subsequent research on harvesting and cultivation.Methods: 1.The samples collected from different areas were combined with the plant morphological characteristics of Paris polyphylla Smith var.chinensis and identified using DNA barcode technology.The main steps included sample preparation,DNA extraction,PCR amplification,sequencing,sequence alignment,species identification,and ITS2 genetic sequence characteristics analysis.2.Paris polyphylla Smith var.chinensis seed samples were collected and examined from seed samples,clarity analysis,germination test,authenticity identification,moisture determination,viability determination,weight measurement,and health measurement.The results were tested for the determination of various indicators in Paris polyphylla Smith var.chinensis seeds.The rationality and focus on the exploration and research on the methods of weight determination and viability testing in the seed standards.The determination of viability was divided into four parts.They were the preparation of test solution of triphenyltetrazolium chloride(TTC),the influencing factors of seed viability testing,and the negative control method,experiment and seed viability testing.3.The stalks were sprayed on a regular basis using paclobutrazol,dildochlor and negative controls.The changes in plant height were measured on a regular basis,and the changes in the color of the Paris polyphylla Smith var.chinensis leaf were observed to determine whether the pesticide used had phytotoxicity to the plant.Other observations included growth,flowering time and seed harvest.4.The new rhizomes,old rhizomes,above-ground parts,and root stalks of Paris polyphylla Smith var.chinensis were collected in the three breeding environments of Shanyin,paddy fields,and under-forest,and the collected parts were measured at different times and different breeding environments.The distribution of saponin content polyphyllins were used as a basis for research on cultivation and harvesting.Results: 1.Among the Paris polyphylla Smith var.chinensis seed identification results,15 samples collected from different producing areas were not completely consistent with those in the GenBank database,and the similarity failed to reach 100%.The results of comparison between the ITS2 sequences of the same basal traits and the accession numbers in the GenBank library were the same,both were GU302308,and the similarity was 100%.2.The seeds of Paris polyphylla Smith var.chinensis seed samples,cleanliness analysis,germination test,authenticity identification,health determination,weight measurement and germination test can be operated according to the standard,but it does not apply to moisture determination.The optimal reaction conditions for the determination of viability was: 0.3% TTC solution concentration for 2.5 to 3 hours at 35°C.3.There was no obvious change in the color of the leaves after spray application.It was proved that the concentration of growth regulator used in the experiment had no obvious injury to the Paris polyphylla Smith var.chinensis seedlings.Compared with the blank group,sprayed paclobutrazol and dillotylamine had a significant effect on slowing plant growth,and the effect of the paclobutrazol is more pronounced than the dihytaamine.4.Paris polyphylla Smith var.chinensis content polyphyllins from high to low distribution: new rhizomes> old rhizomes> aboveground> roots.The contents of new rhizomes and old rhizomes all met the 2015 Chinese Pharmacopoeia standard,and the content of saponins in the new stems was greater than that of old rhizomes,and all parts were mainly polyphyllins VII and polyphyllins VI.Conclusion: 1.The ITS2 sequence has a stable ability to discriminate Paris polyphylla Smith var.chinensis,and it is identified for different leaves shapes,different leaves numbers,and number of sepals.At the same time,it can be seen that the number of leaves,leaves number and leaves shape in different traits of Paris polyphylla Smith var.chinensis should not be used as the basis for their morphological classification.2.In combination with the physiological characteristics of Paris polyphylla Smith var.chinensis seeds,the method for determining the Paris polyphylla Smith var.chinensis seeds' indices was systematically examined,focusing on weight determination,viability testing and other methods,which provided a scientific basis for Paris polyphylla Smith var.chinensis seed quality testing methods.3.The problems of plant lengthiness and easy lodging were initially solved,and research directions were provided to reduce the adverse effects on seed maturity and yield.From the statistical results,it can be seen that although paclobutrazol and dilutedomamine also have the phenomenon of rapid growth of individual plants,the growth rate of the blank control group is significantly higher than that of the normal control group compared with the control group of water spray.Both paclobutrazol and diaminotoluene slowed the growth of the plant,and the paclobutrazol effect was more pronounced.4.Under different breeding conditions,the trend of change in saponin content of Paris polyphylla Smith var.chinensis in various parts of the Paris polyphylla Smith var.chinensis was slightly different,but the contents of root saponins were the highest,and all of them were polyphyllins VII and polyphyllins VI.In different months,there will be a significant increase in the content of polyphyllins,which has a certain reference value for the development of harvest time. |
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