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

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...

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Main Authors: Omar, Siti Ainnsyah, Ashokhan, Sharmilla, Abdul Majid, Nazia, Karsani, Saiful Anuar, Lau, Benjamin Yii Chung, Yaacob, Jamilah Syafawati
Format: Article
Published: Academic Press Inc. 2024
Subjects:
QH301 Biology
T Technology (General)
Online Access:http://eprints.um.edu.my/44833/
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spelling my.um.eprints.448332024-07-01T04:16:45Z http://eprints.um.edu.my/44833/ Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics Omar, Siti Ainnsyah Ashokhan, Sharmilla Abdul Majid, Nazia Karsani, Saiful Anuar Lau, Benjamin Yii Chung Yaacob, Jamilah Syafawati QH301 Biology T Technology (General) 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. Academic Press Inc. 2024 Article PeerReviewed 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 <https://doi.org/10.1016/j.pestbp.2024.105778>. 10.1016/j.pestbp.2024.105778
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic QH301 Biology
T Technology (General)
spellingShingle QH301 Biology
T Technology (General)
Omar, Siti Ainnsyah
Ashokhan, Sharmilla
Abdul Majid, Nazia
Karsani, Saiful Anuar
Lau, Benjamin Yii Chung
Yaacob, Jamilah Syafawati
Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
description 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.
format Article
author Omar, Siti Ainnsyah
Ashokhan, Sharmilla
Abdul Majid, Nazia
Karsani, Saiful Anuar
Lau, Benjamin Yii Chung
Yaacob, Jamilah Syafawati
author_facet Omar, Siti Ainnsyah
Ashokhan, Sharmilla
Abdul Majid, Nazia
Karsani, Saiful Anuar
Lau, Benjamin Yii Chung
Yaacob, Jamilah Syafawati
author_sort Omar, Siti Ainnsyah
title Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
title_short Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
title_full Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
title_fullStr Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
title_full_unstemmed Enhanced azadirachtin production in neem (Azadirachta indica) callus through NaCl elicitation: Insights into differential protein regulation via shotgun proteomics
title_sort enhanced azadirachtin production in neem (azadirachta indica) callus through nacl elicitation: insights into differential protein regulation via shotgun proteomics
publisher Academic Press Inc.
publishDate 2024
url http://eprints.um.edu.my/44833/
_version_ 1805881174047850496
score 13.211869

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