| Pogostemon cablin(Blanco)Benth., a well-known medicinal materials and a kind of flavor, belonging to genus Pogostemon in Lamiaceae. And its dry stem and leaf were both medical parts for clinical use. Herb patchouli was one of aromatic herbs which was pungent-warm in properties, and attribute to lung, spleen and stomach. It was well accepted that herb patchouli could aromatically transform turbid dampness, stop vomiting, release exterior dampness and summer-heat attack. Thus it has been used to remove dampness, relieve summer-heat, exterior syndrome, stop vomiting and stimulate appetite.At the present time, There have been distributions in Hainan province and Guangzhou, Zhaoqing, Zhanjiang in Guangdong province, etc. Commercial herbs of P. cablin were divided into ’Paixiang’(cultivated in Guangzhou),’Zhaoxiang’(cultivated in Zhaoqing),’Zhanxiang’(cultivated in Zhanjiang) and ’Nanxiang’(cultivated in Hainan). In recent years, the demand of P. cablin in domestic and foreign was growing and was in the shortage situation. Varieties were rapidly degraded for consecutive cutting cultivation, which greatly influenced the production of P. cablin. and because ’Paixiang’ and ’Zhaoxiang’ were on the edge of extinction, the indentation of supply and demand was in the augmentation. In addition, degradation of variety had some serious Influence on the stability of P. cablin medication.Therefore germplasm resources of P. cablin could be coordinated and evaluated so as to protect the rare cultivar and exploit resources effectively.Germplasm resources of P. cablin in different planting localities in China were collected in the study. By analysis of genetic diversity of P. cablin at the molecular level, its genetic structure and level of polymorphism were understood. The delineation of the group provided the strong theoretical basis for the development and implementation of source protection of P. cablin. Secondly, the differentiation of rhizogenesis and characteristics of growth and development were compared and analysed. The physio-biochemical changes and ultrastructure characteristic of P. cablin stressed with different concentration of salinity were studied. Finally, there were some studies on quality appraisal of P. cablin from Hainan province.Specific studies as follows:1. The textual researches on a deeper understanding of the origin and the introduction history in our country were carried out in order to clarify the claotic state in the literature and further make good use of its resources.The results showed, the varieties of P. cablin for therapeutic use were not chaotic,while it had produced some chaotic states with Agastache rugosus O.Ktze. in the descriptions of appellation,habitat and characteristic of the plant. It was proved that the term’Huoxiang’in Chinese medical recordations before Ming Dynasty was pointed to P. cablin by the textual researches of materia medica. The cultivated history of/! cablin in our country was traced to Liang Dynasty or before.2. Germplasm resources of P. cablin were utilized based on analysis of RAPD data, the results showed that at species level polymorphic loci percentage (PPL) was78.40%, Nei’S gene diversity index (He)0.2097. At population level, PPL was14.55%-50.23%, averaging at30.16%, Chengyue the highest population, followed by the Zhangpu populations, Hetai, the lowest of populations. Groups of genetic differentiation factor (Gst) was0.4133, population gene flow was0.4466. Data showed that variation came mainly from the variation within populations (58.66%) and among populations(41.34%) although some differentiations but smaller. Cluster analysis revealed that when similar coefficient was0.896,16populations were divided into three material groups.3. Genetic diversity was analysed On the basis of ISSR data. The results showed, at species level, PPL was92.12%, He0.2750. At population level PPL was in35.27%-71.78%, averaging50.21%, Chengyue highest population (71.78%), followed by Yingli (56.85%), Dong’ao the lowest population (35.27%). Gst was0.3396, and Nm was0.9723. The results also showed variation within populations was main source of the total variance (66.07%), and the populations have a certain level of genetic differentiation. Cluster analysis showed when the coefficient was0.886,16populations of P. cablin were divided into four groups.4. SRAP marker was a new technology. It was tested that SRAP marker was applicability in analysis of the genetic diversity of P. cablin. The results showed that at species level, PPL was86.73%, He0.2788. While at populations level of PPL was in35.37%-60.54%, averaging at44.39%. Chengyue, the highest population (60.54%), followed by Damao (55.44%), Dong’ao was the lowest population (35.37%). Gst was to0.4115. Data showed that variation came mainly from the variation within populations (58.86%), the Nm among populations was0.7151, and some differentiations was among populations but smaller. Cluster analysis revealed that when similar coefficient was0.875, 16populations were divided into two groups. By synthetical analyse, the SRAP technology could be used in the studies of genetic structure of P. cablin5. To study the effects of salt stress on the growth and resistant-oxidation enzyme activities of P.cablin, Cutting-rooted seedlings were cultivated in the plastic flowerpot irrigated various concentrations of NaCl (Hoagland solution) for different duration. The results showed, the growth and development of P. cablin was restrained after salt stress. The differentiations of salt stress of lower concentration and little time (less than20mmol·L-1and the9th day) were not obvious. The fresh/dry weight, growth rate and increasing rate of dry matter were obviously decreased with the increasing of NaCl concentration and prolonging of time, but membrane permeability and contents of MD A were increased all along, and lipid peroxidation enhanced, while the root vitality and activities of SOD, POD and CAT were firstly increasing at lower concentration and then decreasing at higher concentration.6. The ultrastructure changes of P. cablin stressed with different concentration of salinity were studied using tranmission electron microscope. The results indicated, the regular and compact chloroplast had integrated membrane system under the water free of salinity; The chloroplast abundant with big starch grain was closed in a mass of mitochondria under low concentration of salinity (40mmol·L-1NaCl); The membrane system of chloroplast departing from cell wall and less of starch grain was degenerated and destroyed, the thylakoids of dissociative chloroplast was intumescent, degenerated and loosened obviously, and the half-vacuolated mitochondria were observed under higher concentration of salinity.7. The characteristics of growth and development were studied in the material of ’Nanxiang’,’Zhanxiang’,’Zhaoxiang’and’Paixiang’. The results showed, the characteristics of rhizogenesis and growth of cutting-rooted seedling had some differentiations. And some effects on P. cablin were come into being under different circumstances of producing area.8. GC method was applied to determine residuals of organochlorine pesticides, the contents of Pb, Cd, Cu, Cr, Hg and As were determined by IPC. Residuals of organochlorine pesticides and heavy metals in soil and P. cablin. from three planting bases in Hainan Province have been analyzed to evaluate the content standard of organochlorine pesticides and heavy metals. The results showed, The contents of residuals of organochlorine pesticides and heavy metals were different among three cultivars and planting bases clearly, but all of the residuals in soil from three planting bases were below the second standard of Environmental quality standard for soils(GB15618), and the residuals in the herbal medicine were also propitious to "Green standards of medicinal plants and preparations for foreign trade and economy"9. Contents and their correlation of mineral nutrient elements (P, K, Ca, Mg, Fe, Zn, Mn, and Cu) in root, stem and leaf of P. cablin. and soil from three planting bases in Hainan Province have been analyzed by ICP to evaluate the content standard of mineral nutrient for instructing the reasonable fertilization. The results showed, the contents of mineral nutrient elements in soil and root, stem and leaf of P. cablin. were different among three planting bases.The order of P, K, Ca, Mg and Mn concentration of root, stem and leaf from high to low was c(leaf)>c(stem)>c(root), and the order of Fe and Cu concentration was c(root)>c(leaf)>c(stem), and the order of Zn concentration was c(root)>c(stem)>c(leaf).A distinctly positive correlation existed between soil and root for K content, as for Zn, there existed a negative correlation.The relationship between Ca and Fe contents in stem and root was significantly positively correlated. P, K, Mg and Cu contents in stem and leaf had significant positive correlation, as for Mn, there was a positive correlation between stem and leaf. As a whole, there was not obvious effects on soil factor in the absorption of mineral nutrient elements.10. The technics conditions of volatile oils extracted by supercritical-CO2fluid extraction (SFE) and steam distillation(SD) were optimized and confirmed. The chemical components and relative contents of essential oil extracted from P. cablin were analyzed and identified by gas chromatography-mass spectrometry combined with computer retrieval respectively. Thirty five and seventeen compounds were identified in the essential oil of leaves and stems obtained by SD, respectively. And the relative contents of exceeding1%had sixteen and twelve compounds, respectively. Patchouli alcohol, the most compound in the essential oil of leaves and stems, was24.32%and33.32%(sheer contents was0.38%and0.23%respectively), respectively. Thirty-nine and twenty-four compounds were identified in the essential oil of leaves and stems obtained by SFE, respectively. And the relative contents of exceeding1%had sixteen and fifteen compounds, respectively. Patchouli alcohol, the most compound in the essential oil of stems, was22.06%(sheer contents0.18%).The major compounds were Azulene, Eudesma, hydroxybenzene, ester, alcohol, aldehyde, ketone, naphthaline, alkyl, acid.At the same time,the quality indexes of medicinal materials of P. cablin in Hainan province were determined.The results established some scientific groundwork for upbuilding quality standard, the fingerprint research, further development of the medicinal material. |
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