Kumar, Ramesh and Kumar, Chavlesh and Choudhury, D R and Ranjan, A and Raipuria, R K and Dubey, K K D and Mishra, A and Kumar, Chetan and Manzoor, M M and Kumar, Ashok and Kumari, A and Singh, K and Singh, G P and Singh, Rakesh (2024) Isolation, Characterization, and Expression Analysis of NAC Transcription Factor from Andrographis paniculata (Burm. f.) Nees and Their Role in Andrographolide Production. Genes (TSI), 15 (4). pp. 1-22. ISSN 2073-4425
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Abstract
Andrographis paniculata (Burm. f.) Nees is an important medicinal plant known for its bioactive compound andrographolide. NAC transcription factors (NAM, ATAF1/2, and CUC2) play a crucial role in secondary metabolite production, stress responses, and plant development through hormonal signaling. In this study, a putative partial transcript of three NAC family genes (ApNAC83, ApNAC21 22 and ApNAC02) was used to isolate full length genes using RACE. Bioinformatics analyses such as protein structure prediction, cis-acting regulatory elements, and gene ontology analysis were performed. Based on in silico predictions, the diterpenoid profiling of the plant’s leaves (five-week-old) and the real-time PCR-based expression analysis of isolated NAC genes under abscisic acid (ABA) treatment were performed. Additionally, the expression analysis of isolated NAC genes under MeJA treatment and transient expression in Nicotiana tabacum was performed. Full-length sequences of three members of the NAC transcription factor family, ApNAC83 (1102 bp), ApNAC21 22 (996 bp), and ApNAC02 (1011 bp), were isolated and subjected to the promoter and gene ontology analysis, which indicated their role in transcriptional regulation, DNA binding, ABA-activated signaling, and stress management. It was observed that ABA treatment leads to a higher accumulation of andrographolide and 14-deoxyandrographolide content, along with the upregulation of ApNAC02 (9.6-fold) and the downregulation of ApNAC83 and ApNAC21 22 in the leaves. With methyl jasmonate treatment, ApNAC21 22 expression decreased, while ApNAC02 increased (1.9-fold), with no significant change being observed in ApNAC83. The transient expression of the isolated NAC genes in a heterologous system (Nicotiana benthamiana) demonstrated their functional transcriptional activity, leading to the upregulation of the NtHMGR gene, which is related to the terpene pathway in tobacco. The expression analysis and heterologous expression of ApNAC21 22 and ApNAC02 indicated their role in andrographolide biosynthesis.
| Item Type: | Article |
|---|---|
| Divisions: | Genebank |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | transcription factor, andrographolide, abscisic acid, medicinal plant, phylogenetic analysis, methyl jasmonate, transient expression |
| Subjects: | Others > Germplasm Others > Gene Bank |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 08 Apr 2025 11:05 |
| Last Modified: | 08 Apr 2025 11:05 |
| URI: | http://oar.icrisat.org/id/eprint/13026 |
| Official URL: | https://www.mdpi.com/2073-4425/15/4/422 |
| Projects: | UNSPECIFIED |
| Funders: | ICAR-National Bureau of Plant Genetic Resources |
| Acknowledgement: | R.K. and R.S. acknowledge the Director, ICAR-NBPGR, New Delhi, for financial support and IARI-National Phytotron Facility New Delhi for providing space for the sowing of plant material, RK is highly thankful to CRP-genomics for providing qRT-PCR thermocycler, Parimalan R (NBPGR, New Delhi), Era V Malhotra (NBPGR, New Delhi), and Rishu Jain (NBPGR, New Delhi) for the help provided in qPCR analysis, Amit Gautam (IIIM, Jammu) and Abhishek Nautiyal (IIIM, Jammu) for the help in biochemical analysis and UGC for Senior Research Fellowship. |
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