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Genome sequencing of herb Tulsi (Ocimum tenuiflorum) unravels key genes behind its strong medicinal properties.

BMC plant biology (2015-09-01)
Atul K Upadhyay, Anita R Chacko, A Gandhimathi, Pritha Ghosh, K Harini, Agnel P Joseph, Adwait G Joshi, Snehal D Karpe, Swati Kaushik, Nagesh Kuravadi, Chandana S Lingu, J Mahita, Ramya Malarini, Sony Malhotra, Manoharan Malini, Oommen K Mathew, Eshita Mutt, Mahantesha Naika, Sathyanarayanan Nitish, Shaik Naseer Pasha, Upadhyayula S Raghavender, Anantharamanan Rajamani, S Shilpa, Prashant N Shingate, Heikham Russiachand Singh, Anshul Sukhwal, Margaret S Sunitha, Manojkumar Sumathi, S Ramaswamy, Malali Gowda, Ramanathan Sowdhamini
ABSTRACT

Krishna Tulsi, a member of Lamiaceae family, is a herb well known for its spiritual, religious and medicinal importance in India. The common name of this plant is 'Tulsi' (or 'Tulasi' or 'Thulasi') and is considered sacred by Hindus. We present the draft genome of Ocimum tenuiflurum L (subtype Krishna Tulsi) in this report. The paired-end and mate-pair sequence libraries were generated for the whole genome sequenced with the Illumina Hiseq 1000, resulting in an assembled genome of 374 Mb, with a genome coverage of 61 % (612 Mb estimated genome size). We have also studied transcriptomes (RNA-Seq) of two subtypes of O. tenuiflorum, Krishna and Rama Tulsi and report the relative expression of genes in both the varieties. The pathways leading to the production of medicinally-important specialized metabolites have been studied in detail, in relation to similar pathways in Arabidopsis thaliana and other plants. Expression levels of anthocyanin biosynthesis-related genes in leaf samples of Krishna Tulsi were observed to be relatively high, explaining the purple colouration of Krishna Tulsi leaves. The expression of six important genes identified from genome data were validated by performing q-RT-PCR in different tissues of five different species, which shows the high extent of urosolic acid-producing genes in young leaves of the Rama subtype. In addition, the presence of eugenol and ursolic acid, implied as potential drugs in the cure of many diseases including cancer was confirmed using mass spectrometry. The availability of the whole genome of O.tenuiflorum and our sequence analysis suggests that small amino acid changes at the functional sites of genes involved in metabolite synthesis pathways confer special medicinal properties to this herb.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Eugenol, ReagentPlus®, 99%
Sigma-Aldrich
1,3-Propanediol, 98%
Sigma-Aldrich
Spectrum Plant Total RNA Kit, sufficient for 50 purifications
Sigma-Aldrich
Ursolic acid, ≥90%
Sigma-Aldrich
GenElute Plant Genomic DNA Miniprep Kit, sufficient for 70 purifications
Sigma-Aldrich
Eugenol, natural, ≥98%, FG
Sigma-Aldrich
Eugenol, ≥98%, FCC, FG