Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics

Omar, Siti Ainnsyah and Ashokhan, Sharmilla and Abdul Majid, Nazia and Karsani, Saiful Anuar and Lau, Benjamin Yii Chung and Yaacob, Jamilah Syafawati (2024) Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics. Pesticide Biochemistry and Physiology, 199. ISSN 0048-3575, DOI https://doi.org/10.1016/j.pestbp.2024.105778.

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Abstract

With their remarkable bioactivity and evolving commercial importance, plant secondary metabolites (PSMs) have gained significant research interest in recent years. Plant tissue culture serves as a credible tool to examine how abiotic stresses modulate the production of PSMs, enabling clear insights into plant stress responses and the prospects for controlled synthesis of bioactive compounds. Azadirachta indica, or neem has been recognized as a repository of secondary metabolites for centuries, particularly for the compound named azadirachtin, due to its bio-pesticidal and high antioxidant properties. Introducing salt stress as an elicitor makes it possible to enhance the synthesis of secondary metabolites, specifically azadirachtin. Thus, in this research, in vitro callus cultures of neem were micro-propagated and induced with salinity stress to explore their effects on the production of azadirachtin and identify potential proteins associated with salinity stress through comparative shotgun proteomics (LCMS/MS). To induce salinity stress, 2-month-old calli were subjected to various concentrations of NaCl (0.05–1.5) for 4 weeks. The results showed that the callus cultures were able to adapt and survive in the salinity treatments, but displayed a reduction in fresh weight as the NaCl concentration increased. Notably, azadirachtin production was significantly enhanced in the salinity treatment compared to control, where 1.5 NaCl-treated calli produced the highest azadirachtin amount (10.847 ± 0.037 mg/g DW). The proteomics analysis showed that key proteins related to primary metabolism, such as defence, energy, cell structure, redox, transcriptional and photosynthesis, were predominantly differentially regulated (36 upregulated and 93 downregulated). While a few proteins were identified as being regulated in secondary metabolism, they were not directly involved in the synthesis of azadirachtin. In conjunction with azadirachtin elicitation, salinity stress treatment could therefore be successfully applied in commercial settings for the controlled synthesis of azadirachtin and other plant-based compounds. Further complementary omics approaches can be employed to enhance molecular-level modifications, to facilitate large-scale production of bioactive compounds in the future. © 2024 Elsevier Inc.

Item Type: Article
Funders: Universiti Malaya, Malaysia [Grant no. ST009-2021], Ministry of Higher Education, Malaysia [Grant no. FP055-2017A, FRGS/1/2017/WAB01/UM/02/2]
Uncontrolled Keywords: Abiotic stress; Azadirachta indica; Azadirachtin; Growth index; Plant tissue culture; Proteomics; Salinity stress
Subjects: Q Science > QH Natural history > QH301 Biology
T Technology > T Technology (General)
Divisions: Faculty of Science > Institute of Biological Sciences
Deputy Vice Chancellor (Research & Innovation) Office > Centre for Research in Biotechnology for Agriculture
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 01 Jul 2024 04:16
Last Modified: 01 Jul 2024 04:16
URI: http://eprints.um.edu.my/id/eprint/44833

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