Systematic identification of chemical forms of key terpene synthase in Cinnamomum camphora / 中国中药杂志

Cinnamomum camphora is an important economic tree species in China. According to the type and content of main components in the volatile oil of leaf, C. camphora were divided into five chemotypes, including borneol-type, camphor-type, linalool-type, cineole-type, and nerolidol-type. Terpene synthase(TPS) is the key enzyme for the formation of these compounds. Although several key enzyme genes have been identified, the biosynthetic pathway of(+)-borneol, which has the most economic value, has not been reported. In this study, nine terpenoid synthase genes CcTPS1-CcTPS9 were cloned through transcriptome analysis of four chemical-type leaves. After the recombinant protein was induced by Escherichia coli, geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) were used as substrates for enzymatic reaction, respectively. Both CcTPS1 and CcTPS9 could catalyze GPP to produce bornyl pyrophosphate, which could be hydrolyzed by phosphohydrolase to obtain(+)-borneol, and the product of(+)-borneol accounted for 0.4% and 89.3%, respectively. Both CcTPS3 and CcTPS6 could catalyze GPP to generate a single product linalool, and CcTPS6 could also react with FPP to generate nerolidol. CcTPS8 reacted with GPP to produce 1,8-cineol(30.71%). Nine terpene synthases produced 9 monoterpene and 6 sesquiterpenes. The study has identified the key enzyme genes responsible for borneol biosynthesis in C. camphora for the first time, laying a foundation for further elucidating the molecular mechanism of chemical type formation and cultivating new varieties of borneol with high yield by using bioengineering technology.

Isolation of Fusarium and identification of its toxins from tuberous root of Pseudostellaria heterophylla / 中国中药杂志

Fusarium is the major pathogen of root rot of Pseudostellaria heterophylla. This study aims to explain the possible distribution of Fusarium species and the contamination of its toxin-chemotypes in tuberous root of P. heterophylla. A total of 89 strains of fungi were isolated from the tuberous root of P. heterophylla. Among them, 29 strains were identified as Fusarium by ITS2 sequence, accounting for 32.5%. They were identified as five species of F. avenaceum, F. tricinctum, F. fujikuroi, F. oxysporum, and F. graminearum based on β-Tubulin and EF-1α genes. LC-MS/MS detected 18, 1, and 5 strains able to produce ZEN, DON, and T2, which accounted for 62.1%, 3.4%, and 17.2%, respectively. Strain JK3-3 can produce ZEN, DON, and T2, while strains BH1-4-1, BH6-5, and BH16-2 can produce ZEN and T2. PCR detected six key synthase genes of Tri1, Tri7, Tri8, Tri13, PKS14, and PKS13 in strain JK3-3, which synthesized three toxins of ZEN, DON, and T2. Four key synthase genes of Tri8, Tri13, PKS14, and PKS13 were detected in strains BH1-4-1, BH6-5, and BH16-2, which were responsible for the synthesis of ZEN and T2. The results showed that the key genes of toxin biosynthesis were highly correlated with the toxins produced by Fusarium, and the biosynthesis of toxin was strictly controlled by the genetic information of the strain. This study provides a data basis for the targeted prevention and control of exo-genous mycotoxins in P. heterophylla and a possibility for the development of PCR for rapid detection of toxin contamination.

Chemical Composition and Antioxidant Activity of Essential Oils from Populations of Baccharis dracunculifolia DC. in Southern Brazil

HIGHLIGHTS Essential oils from populations of B. dracunculifolia were investigated. β-pinene and (E)-nerolidol were the main compounds in B. dracunculifolia populations. The difference in the chemical profile of the essential oils is quantitative only. There is a negative correlation between the antioxidant activity and spathulenol.

Abstract Baccharis dracunculifolia DC. is a Brazilian native plant, presenting wide chemical diversity and numerous pharmaceutical and industrial applications. This research assessed the yield, antioxidant activity and the chemical similarity of essential oils from 10 populations of B. dracunculifolia in the state of Paraná, southern Brazil. The extraction of the volatile compounds was carried out by hydrodistillation, the chemical composition was determined by GC/FID and GC/MS and the antioxidant activity by the DPPH method. The essential oil yield of wild B. dracunculifolia populations ranged from 0.14 to 0.87%. The oils were predominantly composed of oxygenated sesquiterpenes (34.16 - 51.01%), monoterpene hydrocarbons (18.02 - 46.17%) and sesquiterpenes hydrocarbons (9.60 - 17.70%). The major compounds found in all populations were β-pinene (7.65 - 29.8%) and (E)-nerolidol (9.11 - 21.68%). Essential oil solutions (20%) from different populations presented antioxidant capacity ranging from 27.78 to 91.67%. A negative correlation was found between the antioxidant activity and spathulenol (r = -0.696). Multivariate analyses separated the populations into three groups: (1) low concentrations of α-pinene (2.02 - 2.06%), (2) high concentrations of α-pinene (4.17 - 4.61%) and β-pinene (22.54 - 29.80%), and (3) intermediate concentrations of α-pinene (2.38 - 3.31%), β-pinene (12.77 - 19.03%) and spathulenol (6.02 - 9.06%).

Poejo (Cunila galioides Benth. ) Production in Five Agroecological Regions of Rio Grande do Sul

Abstract The objective of this study was to evaluate the biomass and essential oil production of nine populations of poejo (Cunila galioides) cultivated in five agroecological regions of the state of Rio Grande do Sul, under different edaphoclimatic conditions. The experiments were performed in field conditions in Erechim, Caxias do Sul, Pelotas, São Francisco de Paula, and Santa Vitoria do Palmar. The experimental design was completely randomized, with nine populations, eight plants per plot and four repetitions. The following were evaluated: biomass production and essential oil chemical composition and yield. The data underwent ANOVA, followed by Tukey's multiple range test. The adaptability and stability of the populations in the different environments were also evaluated by regression analysis. The results showed great differences between the populations and cultivation sites, with genotype vs. environment interaction. Most populations presented the best biomass production results at Erechim. Pelotas and Santa Vitória do Palmar were the worst locations for poejo production, mainly due to a water deficit occurred during the experiment. The Santa Lucia population presented broad stability and the greatest adaptability to the environments for biomass and essential oil production, but its average production was not satisfactory. The André da Rocha population presented the highest average production of essential oil, and was favored in favorable environments. Regarding essential oil chemical composition, the populations kept stable contents of the major compounds at all locations, with a few variations. In some populations, a higher concentration of sesquiterpenes was observed, which can be attributed to environmental stress.

Quantification of catechin in the spray-dried extract of Pimenta pseudocaryophyllus

Abstract This paper describes the quantification of catechin in the spray-dried extract of Pimenta pseudocaryophyllus (Gomes) Landrum, Myrtaceae, citral chemotype using a validated HPLC-PDA method. The method employs a RP-18 column with acetonitrile:water-orthophosphoric acid 0.05% (gradient system) and UV detection at 210 nm. The method was demonstrated to be simple, sensitive, specific, linear, precise, accurate and robust. The response was linear over a range of 5-200 µg/ml (r > 0.999). The range of recoveries was 92.27-102.54%. The relative standard deviation values for intra- and inter-day precision studies were 4.30 and 3.78%, respectively. This assay can be readily utilized as quality control method for catechin in the dried extract of P. pseudocaryophyllus.

Essential oil from leaves of Myrcianthes fragrans (Myrtaceae) from Costa Rica: a new chemotype? / Aceite esencial de las hojas de Myrcianthes fragrans (Myrtaceae) de Costa Rica: ¿Un nuevo quimiotipo?

Myrcianthes is a Myrtaceous genus of flowering plants of about 30 to 40 species, distributed in the American continent. The aim of this work was to study the chemical composition of the foliar essential oil from M. fragrans growing wild in central Costa Rica. The essential oil was obtained through the steam distillation process in a Clevenger type apparatus. The chemical composition of the oil was performed by capillary gas chromatography with a flame detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS) using the retention indices on a DB-5 type capillary column in addition to mass spectral fragmentation patterns. A total of 98 compounds were identified, accounting for 98.8% of the total amount of the oil. The major constituents in the leaf oil were (E)-methyl cinnamate (39.6%), limonene (34.6%), α-pinene (6.8%), and linalool (6.8%). This is the first report of (E)-methyl cinnamate occurring in oils of this plant genus. These findings appear to suggest a new chemotype of M. fragrans.

Myrcianthes (Myrtaceae) consta de 30 a 40 especies, distribuidas en el continente americano. El objetivo del presente trabajo consistió en identificar la composición química del aceite esencial contenido en las hojas de M. fragrans, planta que crece en forma silvestre en el Valle Central de Costa Rica. La extracción del aceite se efectuó mediante el método de hidrodestilación usando un equipo de Clevenger modificado. La composición química del aceite se analizó mediante las técnicas de cromatografía gaseoso-líquida con detector de ionización de llama (GC-FID) y de cromatografía gaseoso-líquida acoplada a un detector de masas (GC-MS). Se utilizaron índices de retención obtenidos en una columna capilar tipo DB-5 y se compararon con los patrones de iones de fragmentación de masas. Se identificaron en total 98 compuestos, correspondientes a un 98.8% de los constituyentes totales. Los componentes mayoritarios del aceite resultaron ser (E)-cinamato de metilo (39.6%), limoneno (34.6%), α-pineno (6.8%) y linalol (6.8%). Este es el primer informe de la aparición de (E)-cinamato de metilo en aceite de hojas de este género de plantas. Los datos obtenidos parecen sugerir un nuevo quimiotipo de M. fragrans.

Research on effects of chemotype and components of Perilla frutescens leaf volatile oil I: different phenological periods / 中国中药杂志

This experiment researched on three kinds of Perilla frutescens including the widespread PK, PA and rare PL chemotype. The Perilla samples were the mature leaves collected in nutrition, flowering and frutescence three different phenological periods, and at 7 am, 12 pm and 6 pm three day time. The volatile oil was extracted by steam distillationand analyzed by GC-MS, as a result, the three chemotype samples'volatile oil yield was between 0.08% and 0.96%; volatile oil yield of different growth period was as follow: nutrition>flowering>fructescence, and the volatile oil yield of nutrition period: PA type>PK type>PL type. Each chemotype was not affected by the growth and development, indicating that the chemotype is determined by genetic factors. Characteristic and main components of PA and PK type are relatively stable, and the characteristic components of PL type are significantly decreased with the growth. There are still a large number of upstream metabolism components, and the chemical type may have their primitiveness and changeability. The relative content of perillaldehyde, characteristic components of PA type, is basically decreased from morning to night, in all the period. The relative content of perillaketone, characteristic components of PK type, in nutrition and flowering period, when samples were collected at 12 noon is relatively higher than that at 7 am and 6 pm, and contrary to samples collected in frutescence period. The relative content of perillene, characteristic components of PL type, in nutrition and frutescence period are highest at 12 noon, while in flowering period is highest at 6 pm. According to the volatile oil yield and relative content of maincomponents, the best harvest time of PA type is in the morning of the nutrition period; the best harvest time of PK type is in the morning of all the period; and the best harvest time of PL type is at dusk of the nutrition period.

Quimiotipos, Extracción, Composición y Aplicaciones del Aceite Esencial de Lippia alba / Quimiotipos, extração, composição e uso do óleo essencial de Lippia alba / Chemotypes, extraction, chemical composition and use of lippia alba essential oil

RESUMO Lippia alba é uma planta amplamente distribuída nas zonas tropicais, subtropicais e temperadas das Américas, África e Ásia. O óleo essencial de L. alba tem sido amplamente estudado, entretanto apresenta variações de produção. Portanto este estudo teve como objetivo realizar uma revisão dos principais quimiotipos, métodos de extração, composição e aplicação do óleo essencial de L. alba. Neste estudo são discutidos os principais quimiotipos e sua relação com fatores genéticos e características morfológicas. Também são discutidos os fatores que afetam o rendimento de produção, composição química, métodos de extração e do uso e da atividade biológica do óleo essencial de L. alba. Apesar da vasta literatura sobre os óleos essenciais de L. alba, ainda desenvolvimento de aplicações para a produção de cosméticos, fármacos e alimentos, bem como faltam definições agronomicas sobre o cultivo e melhoramento desta planta.

ABSTRACT Lippia alba is a plant widely distributed in tropical, subtropical and temperate zones of the Americas, Africa and Asia. The essential oil of L. alba has been widely studied and there are many variations in the production process. Therefore, this study is aimed at conducting a review of the main chemotypes, extraction methods, composition and application of the essential oil of L. alba. In this study, the main chemotypes and its relation to genetic and morphological characteristics are discussed. It also discusses the factors that affect the yield, chemical composition, extraction methods and the use and the biological activity of the essential oil of L. alba. Despite the vast literature on the essential oils of L. alba, there is still a lack of development in its application for the production of cosmetics, pharmaceuticals and food, as well as a lack of agronomic definitions for its cultivation and genetic improvement.

Atividade antagonista do óleo essencial de Lippia alba (Mill. ) N. E. Brown (Verbenaceae) sobre Meloidogyne incognita (Kofoid & White) Chitwood / Antagonist activity of the essential oil Lippia alba (Mill. ) N. E. Brown (Verbenaceae) on Meloidogyne incognita (Kofoid & White) Chitwood

RESUMO O controle de fitonematóides é uma tarefa difícil. A alta infestação de nematoides no solo obriga os produtores a usarem doses mais elevadas de nematicidas, ou ainda, a aumentarem a freqüência das aplicações, geralmente no solo, ocasionando maior potencial de dano ao homem e ao ambiente. Além disso, nematicidas convencionais vêm sofrendo grandes restrições de uso em muitos países. Desse modo, a busca de novas medidas de controle de fitonematóides é uma prioridade da agricultura sustentável. Desta forma, objetivou-se estudar o efeito de óleos essenciais de Lippia Alba, na mortalidade de juvenis pré-parasitas do segundo estádio de Meloidogyne incognita. O ensaio foi montado em delineamento inteiramente casualizado, com 4 repetições, em esquema fatorial 3×4 (3 quimiotipos x 4 concentrações) empregando-se três diferentes quimiotipos de L. Alba (I, II, III) nas concentrações de 0, 100, 500 e 1000 ppm, coletados em diferentes horas do dia (7, 9, 13, 16 e 19h). Os dados coletados (% de mortalidade) foram submetidos à análise de variância e os valores médios comparados por meio do teste de Tukey. Foi possível concluir que Lippia alba contém, em seu óleo essencial, compostos com efeitos significativos na mortalidade de juvenis de segundo estádio (J2) de M. incognita. Dessa forma, é importante destacar que o acentuado efeito nematicida do óleo essencial de L. alba sugere a possibilidade de seu uso no controle de M. incognita.

ABSTRACT The control of nematodes is difficult. The high infestation of nematodes in the soil requires the producers use higher doses of nematicides, increasing the frequency of applications, usually in soil, causing negative effects to humans and the environment. Furthermore, conventional nematicides have suffered great use restrictions in many countries. Thus, the search for new measures to control nematodes is a priority of sustainable agriculture. Thus, the objective of this study was available the effect of essential oils from Lippia Alba against Meloidogyne incognita. The experiment was conducted in a completely randomized design with four replications in a factorial 3x4 (3 chemotypes and 4 concentrations) using three different chemotypes of L. Alba (I, II, III) at concentrations of 0, 100, 500 and 1000 ppm, collected in different times of day (7 AM, 9 AM, 1 PM, 4 PM e 7 PM). The data collected (% mortality) were subjected to analysis of variance and mean values were compared using the Tukey test. It was able to conclude that L. alba contains in its essential oil, compounds with significant effects on mortality of M. incognita. Thus, it is important to note that the greatest nematicide effect of essential oil of L. alba suggests the possibility of its use to control M. incognita.

Perilla resources of China and essential oil chemotypes of Perilla leaves / 中国中药杂志

This study, based on the findings for Perilla resources, aimed to describe the species, distribution, importance, features, utilization and status of quantitative Perilla resources in China. This not only helps people to know well about the existing resources and researching development, but also indicates the overall distribution, selection and rational use of Perilla resource in the future. According to the output types, Perilla resources are divided into two categories: wild resources and cultivated resources; and based on its common uses, the cultivated resources are further divided into medicine resources, seed-used resources and export resources. The distribution areas of wild resources include Henan, Sichuan, Anhui, Jiangxi, Guangxi, Hunan, Jiangsu and Zhejiang. The distribution areas of medicine resources are concentrated in Hebei, Anhui, Chongqing, Guangxi and Guangdong. Seed-used resources are mainly distributed in Gansu, Heilongjiang, Jilin, Chongqing and Yunnan. Export resource areas are mainly concentrated in coastal cities, such as Zhejiang, Jiangsu, Shandong and Zhejiang. For the further study, the essential oil of leaf samples from different areas were extracted by the steam distillation method and analyzed by GC-MS. The differences in essential oil chemotypes among different Perilla leaves were compared by analyzing their chemical constituents. The main 31 constituents of all samples included: perillaketone (0.93%-96.55%), perillaldehyde (0.10%-61.24%), perillene (52.15%), caryophyllene (3.22%-26.67%), and α-farnesene (2.10%-21.54%). These samples can be classified into following five chemotypes based on the synthesis pathways: PK-type, PA-type, PL-type, PP-type and EK-type. The chemotypes of wild resources included PK-type and PA-type, with PK-type as the majority. All of the five chemotypes are included in cultivated resources, with PA-type as the majority. Seed-used resources are all PK-type, and export resources are all PA-type. The P. frutescens var. frutescens include five chemotypes, with PK-type as the majority. The PK-type leaves of P. frutescens var. acuta are green, while the PA-type leaves are reddish purple. The P. fruteseens var. crispa was mainly PA type with reddish purple leaves. The differences of the main chemotypes provide a scientific basis for distinguishing between Zisu and Baisu in previous literatures. Based on the lung toxicity of PK and the traditional use of Perilla, the testing standard of essential oil and Perilla herb shall be built, and PA type is recommended to be used in traditional Chinese medicine.

Analysis on Chemotype of Volatile Oil of Mentha crispata Schrad. ex Willd / 世界科学技术-中医药现代化

This study was aimed to analyze the volatile oil of Mentha crispata Schrad. ex Willd. in order to provide evidence for its chemotype and guidance for its production application. The chemical analysis was detected by headspace GC-MS. The results showed that 64 chemical compounds were detected. It was concluded that the volatile oil of M. crispata Schrad. ex Willd. mainly contained eucalyptol (35.58%), limonene (16.92%) and pinene (15.33%). It was concluded that the analysis on composition characteristics and main compounds of M. crispata Schrad. ex Willd. can provide evidences in its production application and chemotype.

Phytochemical composition of the essential oil of different populations of Stachys lavandulifolia Vahl

Objective: To examine the chemical variability in inflorescences of wild populations of Stachys lavandulifolia Vahl (S. lavandulifolia) collected throughout two provinces (Isfahan and Chaharmahal va Bakhtiary), Southwest Iran. Methods: The essential oils of S. lavandulifolia Vahl from seven locations were obtained by hydro-distillation and analysed by gas chromatography and gas chromatography-mass spectrometry. Results: The results revealed that distinct differences in the content of compounds depending on region of sample collection. The main constituents of the essential oils were α-thujone (0.3%-32.3%), α-pinene (trace to 37.3%), myrcene (0.5%-15.9%),β-phellandrene (1.1%-37.9%), germacrene D (0.4%-11.3%),?-cadinene (trace to 11.6%) and 1, 4-methano-1 H-indene (trace to 10.1%). Conclusions:The results of the present study indicated that essential oil components of S. lavandulifolia Vahl can be varied with genetic (ecotype), environmental conditions and geographic origin. In general, the essential oils of various populations of S. lavandulifolia Vahl were rich in monoterpenoids and sesquiterpenoids.

Phytochemical composition of the essential oil of different populations of Stachys lavandulifolia Vahl

<p><b>OBJECTIVE</b>To examine the chemical variability in inflorescences of wild populations of Stachys lavandulifolia Vahl (S. lavandulifolia) collected throughout two provinces (Isfahan and Chaharmahal va Bakhtiary), Southwest Iran.</p><p><b>METHODS</b>The essential oils of S. lavandulifolia Vahl from seven locations were obtained by hydro-distillation and analysed by gas chromatography and gas chromatography-mass spectrometry.</p><p><b>RESULTS</b>The results revealed that distinct differences in the content of compounds depending on region of sample collection. The main constituents of the essential oils were α-thujone (0.3%-32.3%), α-pinene (trace to 37.3%), myrcene (0.5%-15.9%), β-phellandrene (1.1%-37.9%), germacrene D (0.4%-11.3%), Δ-cadinene (trace to 11.6%) and 1, 4-methano-1 H-indene (trace to 10.1%).</p><p><b>CONCLUSIONS</b>The results of the present study indicated that essential oil components of S. lavandulifolia Vahl can be varied with genetic (ecotype), environmental conditions and geographic origin. In general, the essential oils of various populations of S. lavandulifolia Vahl were rich in monoterpenoids and sesquiterpenoids.</p>