Genetic Diversity Analysis of Pogostemon cablin Based on AFLP Markers / 中国实验方剂学杂志

Objective:This paper aims to study the genetic diversity of <italic>Pogostemon cablin</italic> by amplified fragment length polymorphism （AFLP） markers. Method:The 12 pairs of primers were used for AFLP analysis of 212 samples from 14 varieties，and biological analysis software such as POPGENE 32，Arlequinver 3.5，MEGA 7 and NTSYSpc 2.10e were used for polymorphism parameter calculation，principal coordinate analysis and cluster analysis. Result:A total of 2 238 loci were amplified by 12 pairs of primers. 2 226 of them were polymorphic loci， accounting for 99.38%. At the inter-population level，the values of effective alleles（<italic>Ne</italic>），Nei's gene diversity index（<italic>H</italic>），Shannon polymorphic information index（<italic>I</italic>） were 1.365 6±0.066 3， 0.220 7±0.036 4， and 0.343 7±0.050 2，respectively；and 1.118 5±0.038 7，0.071 3±0.023 0，0.109 4±0.035 0，respectively at the intra-population level. Analysis of molecular variance（AMOVA）showed that 71.57% of the total variation of <italic>P. cablin</italic> was of inter-population nature， and 28.43% was of intra-population nature. The 14 populations could be divided into four groups by cluster analysis. Conclusion:The results of AFLP molecular markers showed that abundant genetic diversity was present at inter-population level of <italic>P. cablin</italic>，however，relatively low at intra-population level； the genetic differentiation at the inter-population level was significant，which could provide a reference for the subsequent study of good germplasm selection of <italic>P. cablin</italic>.

Effects of 5-azacytidine on DNA methylation and index components in patchouliol-type Pogostemon cablin / 中国中药杂志

This study aims to explore the relationship of DNA methylation with the contents of the index components as well as the growth and development of Pogostemon cablin. The demethylation reagent 5-azacytidine(5-azaC) was used to treat the tissue culture seedlings of patchouliol-type P. cablin. High performance liquid chromatography was employed to evaluate the changes of DNA methy-lation in P. cablin, and GC-MS to detect the contents of index components in P.cablin. The agronomic characters of P.cablin were measured using the common methods. The results showcased that DNA methylation of P.cablin was significantly reduced by 5-azaC in a concentration-dependent manner. Thirty days after treatment with 5-azaC at different concentrations, the content of patchouli alcohol changed slightly; compared with that in the control group, the content of pogostone in 50 μmol·L~(-1) and 100 μmol·L~(-1) 5-azaC groups was significantly up-regulated. The 100 μmol·L~(-1) 5-azaC group had the largest differences in contents of pogostone and patchouli alcohol compared with the control group, followed by the 50 μmol·L~(-1) 5-azaC group. Ninety days after disinhibition, the content of pogostone in the treatment group was significantly increased and the content of patchouli alcohol was significantly decreased. In addition, 5-azaC significantly inhibited the growth and development of P.cablin in a dose-dependent manner. These results indicate that DNA methylation regulates the biosynthesis of the index components in patchouliol-type P.cablin and proper demethylation can directly promote the synthesis of pogostone and indirectly affect the accumulation of patchouli alcohol.

Biosynthesis and metabolism regulation of terpenoids in Pogostemon cablin: a review / 中国中药杂志

The terpenoids in Pogostemon cablin have complex structures and abundant pharmacological effects. Patchouli alcohol(PA) and pogostone(PO) have a high medicinal value by virtue of anti-tumor, anti-inflammatory, antibacterial, antioxidant, and other biological activities. Due to the low content of terpenoid metabolites in P. cablin, the study of biosynthesis and metabolism regulation can provide a biosynthetic basis for obtaining high-content terpenoids. In this study, key enzyme genes in biosynthesis, transcription factors in metabolism regulation, spatio-temporal expression of terpene synthase were reviewed, aiming to provide a reference for the development, protection, and utilization of P. cablin resources.

Study on cytotoxic secondary metabolites of endophytic fungus Diaporthe longicolla A616 from Pogostemon cablin / 中国中药杂志

To study active secondary metabolites of endophytic fungus Diaporthe longicolla A616 isolated from Pogostemon cablin. Ten compounds were isolated from fermentation product of the strain 616 by silica gel, reverse phase silica gel, Sephadex-LH20, HPLC and so on. Their structures were identified as 1,3-diamino-1,3-dimethylurea(1),(7R,9R)-7-hydroxy-9-propyl-5-nonen-9-olide(2), Ergosta-5,7,22-trien-3β-ol(3),(22E,24R)-ergosta-4,6,8(14)-22-tetraen-3-one(4),(22E,24R)-3β,5α-dihydroxy-6β-ergosta-7,22-diene(5), citreoisocoumarin(6), glycerol monolinoleate(7), 1-(2-hydroxyethoxy)ethyl(E)-octadec-9-enoate(8), cyclo-(L-Pro-L-Ala)(9), cyclo(L)-Pro-(L)-Val(10), respectively, based on extensive spectroscopic analysis and literature comparisons. Compounds 6-10 were isolated from the genus Diaporthe for the first time. All isolated compounds were evaluated for in vitro cytotoxic activities against SF-268, MCF-7, NCI-H460 and HepG-2 tumor cell lines. Compounds 4 and 5 showed potent growth inhibitory activities against the four cell lines with IC₅₀ values of 5.3, 6.5, 12.2, 6.1μmol•L⁻¹ and 8.2, 5.2, 6.1, 9.4μmol•L⁻¹, respectively.

Cultural regionalization for Notopterygium incisum based on 3S technology platform. I. Evaluation for growth suitability for N. incisum based on ecological factors analysis by Maxent and ArcGIS model / 中国中药杂志

Growth suitability as assessment indicators for medicinal plants cultivation was proposed based on chemical quality determination and ecological factors analysis by Maxent and ArcGIS model. Notopterygium incisum, an endangered Chinese medicinal plant, was analyzed as a case, its potential distribution areas at different suitability grade and regionalization map were formulated based on growth suitability theory. The results showed that the most suitable habitats is Sichuan province, and more than 60% of the most suitable areawas located in the western Sichuan such as Aba and Ganzi prefectures for N. incisum. The results indicated that habitat altitude, average air temperature in September, and vegetation types were the dominant factors contributing to the grade of plant growth, precipitation and slope were the major factors contributing to notopterol accumulation in its underground parts, while isoimperatorin in its underground parts was negatively corelated with precipitation and slope of its habitat. However, slope as a factor influencing chemical components seemed to be a pseudo corelationship. Therefore, there were distinguishing differences between growth suitability and quality suitability for medicinal plants, which was helpful to further research and practice of cultivation regionalization, wild resource monitoring and large-scale cultivation of traditional Chinese medicine plants.

Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis / 中国中药杂志

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

Study on determination and principal component analysis of inorganic elements in Polygonum multiflorum from different areas / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the character of inorganic elements in Polygonum multiflorum.</p><p><b>METHOD</b>The contents of elements such as Al, B, Ba, Ca, Cu, Mn, Mg, Fe, Na, Ni, P, Se, Sr, Ti and Zn in nine P. multiflorum samples were determined by means of ICP-AEC. The results were used for the development of element distrubution diagram. The principal component analysis and one-way ANOVA of SPSS were applied for the study of characteristic elements in P. multiflorum.</p><p><b>RESULT</b>The contents of Al, Ca, K, Mg, Sr,Ti in wild P. multiflorum were remarkable higher than those in cultured P. multiflorum, and there was no significant difference between cultured and wild in the other elements. Five principal components which accounted for over 90% of the total variance were extracted from the original data. The analysis results show that Al, B, Ba, Fe, Na, Ni, Ti, Ca and Sr may be the characteristic elements in P. multiflorum. The element distrubution diagram of the sample from Tianyang was remarkable different comparing with the others.</p><p><b>CONCLUSION</b>The principal component analysis could be used in data processing in inorganic elements.</p>