Abstract
BackgroundBacopa monnieri commonly known as Brahmi is utilized in Ayurveda to improve memory and many other human health benefits. Bacosides enriched standardized extract of Bacopa monnieri is being marketed as a memory enhancing agent. In spite of its well known pharmacological properties it is not much studied in terms of transcripts involved in biosynthetic pathway and its regulation that controls the secondary metabolic pathway in this plant. The aim of this study was to identify the potential transcripts and provide a framework of identified transcripts involved in bacosides production through transcriptome assembly.ResultsWe performed comparative transcriptome analysis of shoot and root tissue of Bacopa monnieri in two independent biological replicate and obtained 22.48 million and 22.0 million high quality processed reads in shoot and root respectively. After de novo assembly and quantitative assessment total 26,412 genes got annotated in root and 18,500 genes annotated in shoot sample. Quality of raw reads was determined by using SeqQC-V2.2. Assembled sequences were annotated using BLASTX against public database such as NR or UniProt. Searching against the KEGG pathway database indicated that 37,918 unigenes from root and 35,130 unigenes from shoot were mapped to 133 KEGG pathways. Based on the DGE data we found that most of the transcript related to CYP450s and UDP-glucosyltransferases were specifically upregulated in shoot tissue as compared to root tissue. Finally, we have selected 43 transcripts related to secondary metabolism including transcription factor families which are differentially expressed in shoot and root tissues were validated by qRT-PCR and their expression level were monitored after MeJA treatment and wounding for 1, 3 and 5 h.ConclusionsThis study not only represents the first de novo transcriptome analysis of Bacopa monnieri but also provides information about the identification, expression and differential tissues specific distribution of transcripts related to triterpenoid sapogenin which is one of the most important pharmacologically active secondary metabolite present in Bacopa monnieri. The identified transcripts in this study will establish a foundation for future studies related to carrying out the metabolic engineering for increasing the bacosides biosynthesis and its regulation for human health benefits.
Highlights
Bacopa monnieri commonly known as Brahmi is utilized in Ayurveda to improve memory and many other human health benefits
Transcriptome sequencing and de novo assembly To characterize the transcriptome of Bacopa monnieri, we sequenced cDNA libraries prepared from the shoot and root tissues of Bacopa monnieri using Illumina HI Seq 2000 System yielding an entire 24.85 million and 24.31 million raw reads
Total 22.48 and 22.0 million reads were generated after sequencing of the shoot and root tissues of Bacopa monnieri subsequently after removing adapter containing low quality reads
Summary
Bacopa monnieri commonly known as Brahmi is utilized in Ayurveda to improve memory and many other human health benefits. It has been used worldwide due to its broad use of pharmaceutically important triterpenoid saponins mainly bacosides [1] It is a perennial, creeping herb basically found in the wetlands of Australia, southern India, Europe, Asia, Africa, and South America. In the conventional system of Indian medicine, Bacopa is well treated as a drug to augment intelligence and memory function and combat the effects of mental stress [2]. It is used for gastrointestinal infections, rejuvenation, skin disorders, epilepsy, pyrexia and analgesia [3]. Earlier studies revealed that bacosides from Bacopa monnieri leaf extract is used to treat memory functions and impairment [6]. Evaluation and extraction of bacosides from areal part of the plant using HPLC chromatographic technique have been performed and reported that highest concentration of bacoside A is present in stolon (9.54 mg/g dry wt) followed by leaves (4.73 mg/g dry wt) and roots [7]
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