Transcriptome Analysis of Buds and Leaves Using 454 Pyrosequencing to Discover Genes Associated with the Biosynthesis of Active Ingredients in Lonicera japonica Thunb.

Abstract

Background Lonicera japonica Thunb. is a plant used in traditional Chinese medicine known for its anti-inflammatory, anti-oxidative, anti-carcinogenic, and antiviral pharmacological properties. The major active secondary metabolites of this plant are chlorogenic acid (CGA) and luteoloside. While the biosynthetic pathways of these metabolites are relatively well known, the genetic information available for this species, especially the biosynthetic pathways of its active ingredients, is limited.Methodology/Principal FindingsWe obtained one million reads (average length of 400 bp) in a whole sequence run using a Roche/454 GS FLX titanium platform. Altogether, 85.69% of the unigenes covering the entire life cycle of the plant were annotated and 325 unigenes were assigned to secondary metabolic pathways. Moreover, 2039 unigenes were predicted as transcription factors. Nearly all of the possible enzymes involved in the biosynthesis of CGA and luteoloside were discovered in L. japonica. Three hydroxycinnamoyl transferase genes, including two hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase genes and one hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT) gene featuring high similarity to known genes from other species, were cloned. The HCT gene was discovered for the first time in L. japonica. In addition, 188 candidate cytochrome P450 unigenes and 245 glycosyltransferase unigenes were found in the expressed sequence tag (EST) dataset.ConclusionThis study provides a high quality EST database for L. japonica by 454 pyrosequencing. Based on the EST annotation, a set of putative genes involved in CGA and luteoloside biosynthetic pathways were discovered. The database serves as an important source of public information on genetic markers, gene expression, genomics, and functional genomics in L. japonica.

De novo transcriptome assembly and characterization of nine tissues of Lonicera japonica to identify potential candidate genes involved in chlorogenic acid, luteolosides, and secoiridoid biosynthesis pathways

“…Lonicera japonica Thunb, also known as Japanese honeysuckle, 'Jin Yin Hua', and 'Ren Dong', belongs to the Caprifoliaceae family and is often used in traditional Chinese and Japanese medicine [1]. L. japonica is native to eastern Asia, and is cultivated worldwide, particularly in China, Japan, and Korea due to its medicinal properties, and as an ornamental plant due to its pleasant smelling flowers, and attractive evergreen foliage [2]. However, as it is highly invasive to the ecology of some countries, such as New Zealand and several other countries including North America, it is considered a major nuisance and is restricted [3].…”

“…L. japonica has been used as traditional medicine in China for over thousands of years, and has been listed as top grade in 'Ming Yi Bie Lu' and 'Shen Nong Ben Cao Jing', and described in 'Ben Cao Gang Mu', the famous classical book of Chinese Materia Medica, as early as the seventeenth century, for applications in various diseases such as to clean away the heat-evil or heal the swelling [1]. Different parts of L. japonica have been reported to possess unique medicinal properties, with flowers and floral buds being highly used in Chinese traditional medicine, while the leaves and stems are used in Japan [2,4,5]. The commercial value of L. japonica in the herbal medicine trading market has increased by several hundred-fold in recent years, and [30 % of current traditional Chinese medicine prescriptions contain extracts from different plant parts of L. japonica [6].…”

“…Whole plant or aerial parts of L. japonica, particularly leaves and floral buds are used to derive bioactive metabolites for various preparations and medicinal uses [7]. Modern pharmacological studies have shown that extracts from L. japonica possess a wide range of bioactive properties, such as anti-bacterial, anti-inflammatory, antiviral, anti-pyretic, anti-oxidant, anti-hyperlipidemic, and anti-nociceptive among others [2,5,[8][9][10][11][12][13][14][15]. Extracts from L. japonica were used to prevent and treat severe acute respiratory syndromes, H1N1 influenza, and hand, foot and mouth diseases, and were reported to be effective against SARS coronavirus [2].…”

Chlorogenic acid inhibits cholestatic liver injury induced by α-naphthylisothiocyanate: involvement of STAT3 and NFκB signalling regulation

“…At present, traditional phytotherapy and plant‐derived natural medicine are quite popular for treating liver diseases. Lonicera japonica Thunb (Caprifoliaceae family), known as Jinyinhua (in Chinese), is widely consumed as healthy tea beverage and used in traditional Chinese medicine (TCM) to treat various diseases including hepatitis, arthritis, allergy and infection . Of particular interest is the L. japonica ‐containing TCM formula: Yin‐zhi‐huang (YZH) is a classical polyherbal prescription dated from the traditional Chinese prescription ‘Yin‐chen‐hao‐tang’, which has been widely used in East Asian countries for more than a thousand years to treat jaundice and chronic liver diseases .…”

“…Of particular interest is the L. japonica ‐containing TCM formula: Yin‐zhi‐huang (YZH) is a classical polyherbal prescription dated from the traditional Chinese prescription ‘Yin‐chen‐hao‐tang’, which has been widely used in East Asian countries for more than a thousand years to treat jaundice and chronic liver diseases . Extensive studies on the phytochemical ingredients from coffee and L. japonica demonstrated that chlorogenic acid (CGA) is supposed to be the primary active component (indicator compound) contributing to the health benefits …”