PatJAZ6 Acts as a Repressor Regulating JA-Induced Biosynthesis of Patchouli Alcohol in Pogostemon Cablin.

The JASMONATE ZIM DOMAIN (JAZ) proteins act as negative regulators in the jasmonic acid (JA) signaling pathways of plants, and these proteins have been reported to play key roles in plant secondary metabolism mediated by JA. In this study, we firstly isolated one JAZ from P. cablin, PatJAZ6, which was characterized and revealed based on multiple alignments and a phylogenic tree analysis. The result of subcellular localization indicated that the PatJAZ6 protein was located in the nucleus of plant protoplasts. The expression level of PatJAZ6 was significantly induced by the methyl jasmonate (MeJA). Furthermore, by means of yeast two-hybrid screening, we identified two transcription factors that interact with the PatJAZ6, the PatMYC2b1 and PatMYC2b2. Virus-induced gene silencing (VIGS) of PatJAZ6 caused a decrease in expression abundance, resulting in a significant increase in the accumulation of patchouli alcohol. Moreover, we overexpressed PatJAZ6 in P. cablin, which down-regulated the patchoulol synthase expression, and then suppressed the biosynthesis of patchouli alcohol. The results demonstrate that PatJAZ6 probably acts as a repressor in the regulation of patchouli alcohol biosynthesis, contributed to a model proposed for the potential JA signaling pathway in P. cablin.

Phylogenetic relationships, character evolution and biogeographic diversification of Pogostemon s.l. (Lamiaceae).

Pogostemon (Lamiaceae; Lamioideae) sensu lato is a large genus consisting of about 80 species with a disjunct African/Asian distribution. The infrageneric taxonomy of the genus has historically been troublesome due to morphological variability and putative convergent evolution within the genus. Notably, some species of Pogostemon are obligately aquatic, perhaps the only Lamiaceae taxa which exhibit this trait. Phylogenetic analyses using the nuclear ribosomal internal transcribed spacer (ITS) and five plastid regions (matK, rbcL, rps16, trnH-psbA, trnL-F), confirmed the monophyly of Pogostemon and its sister relationship with the genus Anisomeles. Pogostemon was resolved into two major clades, and none of the three morphologically defined subgenera of Pogostemon were supported as monophyletic. Inflorescence type (spikes with more than two lateral branches vs. a single terminal spike, or rarely with two lateral branches) is phylogenetically informative and consistent with the two main clades we recovered. Accordingly, a new infrageneric classification of Pogostemon consisting of two subgenera is proposed. Molecular dating and biogeographic diversification analyses suggest that Pogostemon split from its sister genus in southern and southeast Asia in the early Miocene. The early strengthening of the Asia monsoon system that was triggered by the uplifting of the Qinghai-Tibetan Plateau may have played an important role in the subsequent diversification of the genus. In addition, our results suggest that transoceanic long-distance dispersal of Pogostemon from Asia to Africa occurred at least twice, once in the late Miocene and again during the late-Miocene/early-Pliocene.

The Complete Chloroplast Genome Sequences of the Medicinal Plant Pogostemon cablin.

Pogostemon cablin, the natural source of patchouli alcohol, is an important herb in the Lamiaceae family. Here, we present the entire chloroplast genome of P. cablin. This genome, with 38.24% GC content, is 152,460 bp in length. The genome presents a typical quadripartite structure with two inverted repeats (each 25,417 bp in length), separated by one small and one large single-copy region (17,652 and 83,974 bp in length, respectively). The chloroplast genome encodes 127 genes, of which 107 genes are single-copy, including 79 protein-coding genes, four rRNA genes, and 24 tRNA genes. The genome structure, GC content, and codon usage of this chloroplast genome are similar to those of other species in the family, except that it encodes less protein-coding genes and tRNA genes. Phylogenetic analysis reveals that P. cablin diverged from the Scutellarioideae clade about 29.45 million years ago (Mya). Furthermore, most of the simple sequence repeats (SSRs) are short polyadenine or polythymine repeats that contribute to high AT content in the chloroplast genome. Complete sequences and annotation of P. cablin chloroplast genome will facilitate phylogenic, population and genetic engineering research investigations involving this particular species.

Combined fluorescence-transmittance imaging system for geographical authentication of patchouli oil.

Recently, demand for authentication technology is growing rapidly in an attempt to overcome counterfeiting of high-value agricultural products, such as patchouli oil. Fingerprinting methods based on spectroscopy are one such technology being used for authentication. However, the spectral datasets obtained are multivariate in nature; containing thousands of data points for a single sample, making data acquisition and processing time-consuming. Therefore, reduction and simplification in the number of variables used required is needed to provide a more rapid and applicable method. Color cameras, which can capture image in the visible region light, could be such an alternative spectral data acquisition approach. In this research, a simplified spectroscopy method was developed for origin authentication of patchouli oil. The system consists of front ultraviolet light induced (365 nm) fluorescence and a white LED-based backlighting imaging system that consecutively captures the fluorescence and transmittance characteristics of the oil in the visible region. From the captured images, features were extracted and analyzed using Principle Component Analysis (PCA) to identify important image features for discrimination of origin. From the samples measured, the samples clustered around three islands of origin in the PCA space. A classification model based on fluorescence and transmittance image features (color values) could discriminate origin classes with a total accuracy of 88.46%. A lower accuracy was found for the Java class due to low sample numbers. This result demonstrates that the proposed system has the potential to be a rapid authentication tool for determining the geographical origin of patchouli oils.

Rendimento e composição do óleo essencial de patchouli (Pogostemon cablin) conforme o tempo de extração / Yield and composition of the patchouli (Pogostemon cablin) essential oil according to the extraction time

O patchouli possui óleo essencial nas folhas com utilização principalmente na indústria de perfumaria. O objetivo foi avaliar o melhor tempo de extração de óleo essencial de folhas secas de patchouli. Os tratamentos foram 1, 2, 3, 4, 5, 6, 7 e 8 horas de extração, através do método de hidrodestilação, com aparelho graduado do tipo Clevenger e balões com capacidade de 2 L.O delineamento foi inteiramente casualizado com três repetições. O material destilado foi seco à sombra até atingir aproximadamente 20% de umidade. Para cada tratamento, foram utilizadas amostras de 50 g de massa seca foliar. Foram avaliados o rendimento e a composição do óleo essencial. Não houve diferença entre os diferentes tempos de extração no rendimento de óleo essencial, podendo a extração do óleo essencial de patchouli ser realizada com uma hora de extração. O tempo de extração aumenta as porcentagens relativas do beta-guaieno (0,81%), beta-patchouleno (1,26%), alfa-selineno (1,37%), cariofileno (2,44%), alfa-patchouleno (3,08%) e gama-patchouleno (4,82%). O teor de pogostol (5,11%) reduz com o aumento do tempo de extração. O patchoulol, alfa-guaieno, alfa-bulneseno e seicheleno não sofrem influencia do tempo de extração.

Patchouli accumulatea an essential oil on its leaves, and it is mainly used by the perfume industry. The purpose of this work was to determine the distillation time for the extraction of essential oils from the leaves of patchouli. The treatments included 1, 2, 3, 4, 5, 6, 7 and 8 hours of extraction through the hydrodistillation method, using a Clevenger apparatus. The experimental design was completely randomized with three replications. The leaves were dried at room temperature until they reached 20% of humidity. The essential oil yield was measured from samples containing 50g of leaf dry mass. The essential oil production and composition were evaluated. No differences among the treatments were found on the essential oil yield, suggesting that the essential oil extraction can be carried out for 1 hour according to the experimental conditions. However, extending the extraction time, an increase in the relative percentage of beta-patchoulene (1.26%), cariofilene (2.44%), gamma-patchoulene (4.82%), alpha-patchoulene (3.08%), beta-guaiene (0.81%) and alpha-selinene (1.37%) was observed. The pogostol (5.11%) content was reduced with the increase of the extraction time. Patchoulol, alpha-guaieno, alpha-bulneseno and seicheleno contents were not affected by the evaluated extraction times.