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6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize

Aminochitosan, derived from chitosan, features enhanced water solubility, and improved antifungal efficacy attributed to an additional amine group at the C-6 position, hinting at its superior antifungal potential compared to native chitosan. This PhD thesis aimed to explore the optimal concentration...

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Main Author: Moola, Naadirah
Other Authors: Rafudeen, Mohamed
Format: Thesis
Language:Eng
Published: Department of Molecular and Cell Biology 2025
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access_status_str Open Access
author Moola, Naadirah
author2 Rafudeen, Mohamed
author_browse Moola, Naadirah
Rafudeen, Mohamed
author_facet Rafudeen, Mohamed
Moola, Naadirah
author_sort Moola, Naadirah
collection Thesis
description Aminochitosan, derived from chitosan, features enhanced water solubility, and improved antifungal efficacy attributed to an additional amine group at the C-6 position, hinting at its superior antifungal potential compared to native chitosan. This PhD thesis aimed to explore the optimal concentration of aminochitosan and its molecular weight fractions for enhanced antifungal and priming properties in the tomato/Botrytis cinerea and maize/ Fusarium verticillioides pathosystems. In the B. cinerea pathosystem, various concentrations of aminochitosan were assessed for their antifungal effectiveness against B. cinerea growth and sporulation. Additionally, in planta studies were conducted to phenotype and quantify temporal stress responses using both qualitative and quantitative chlorophyll fluorescence imaging as well as DAB assays combined with microscopy. Despite batch-to-batch variations, aminochitosan consistently inhibited fungal growth and sporulation in a dose-dependent manner. In planta, aminochitosan pre-treatment induced robust defence responses in tomato leaves, resulting in a resistant phenotype that was mediated through a combination of enhanced photosynthetic efficiency as evidenced by enhanced Fv/Fm and chlorophyll content. The DAB assays suggested that these resistant phenotypes were also ROS-independent (H2O2 specifically) due to the strong positive impact of direct inhibition. The resistant phenotype and optimal efficacy of the aminochitosan MW fractions was observed at 3.5-15 kDa for antifungal efficacy and 15-20 kDa for in planta efficacy. Consequently, leaf senescence, hypersensitive responses and therefore necrosis were mitigated suggesting that aminochitosan primed defence responses in both mock and B. cinerea inoculated leaves. A temporal, label-free quantitative proteomic analysis revealed the differential priming of key molecular mechanisms underpinning aminochitosan primed states both with and without B. cinerea infection at 6 and 9 hpi in the tomato/B. cinerea pathosystem. Aminochitosan treatment (1 mg/mL) differentially regulated proteins as early as 6 hpi with some of the induced responses being sustained up to 9 hpi. Additionally, several proteins were oppositely regulated between aminochitosan pre-treatment and B. cinerea infection, indicating differential regulation patterns between the “primed state” and the “triggered state”. The proteomic data therefore partially validated the ‘priming' capacity of aminochitosan in 5- week-old tomato leaves, specifically diamino 3 when used as a foliar spray at a concentration of 1 mg/mL. The bimodal effects of aminochitosan in a maize/Fusarium verticillioides pathosystem were investigated by assessing the direct antifungal efficacy and elicitation of plant defence properties in planta. Aminochitosan displayed significant antifungal activity on both radial growth and sporulation at a minimum concentration of 1 mg/mL. Aminochitosan was also assessed in planta as preventative or curative treatments in maize seedlings infected with F. verticillioides at two time points. In the preventative treatment, salicylic acid accumulated during the early stages of infection (biotrophic phase) whereas in the curative treatment, jasmonic acid accumulated during the necrotrophic phase. In summary, we demonstrated that water-soluble aminochitosan possesses key properties that enable plant priming in addition to its superior direct antifungal activity compared to native chitosan. We also identified the optimal molecular weights and concentrations necessary for achieving maximum and 50% inhibition activity
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license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
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spelling oai:open.uct.ac.za:11427/41146 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize Moola, Naadirah Rafudeen, Mohamed Jardine Mogamat Molecular and Cell Biology Aminochitosan, derived from chitosan, features enhanced water solubility, and improved antifungal efficacy attributed to an additional amine group at the C-6 position, hinting at its superior antifungal potential compared to native chitosan. This PhD thesis aimed to explore the optimal concentration of aminochitosan and its molecular weight fractions for enhanced antifungal and priming properties in the tomato/Botrytis cinerea and maize/ Fusarium verticillioides pathosystems. In the B. cinerea pathosystem, various concentrations of aminochitosan were assessed for their antifungal effectiveness against B. cinerea growth and sporulation. Additionally, in planta studies were conducted to phenotype and quantify temporal stress responses using both qualitative and quantitative chlorophyll fluorescence imaging as well as DAB assays combined with microscopy. Despite batch-to-batch variations, aminochitosan consistently inhibited fungal growth and sporulation in a dose-dependent manner. In planta, aminochitosan pre-treatment induced robust defence responses in tomato leaves, resulting in a resistant phenotype that was mediated through a combination of enhanced photosynthetic efficiency as evidenced by enhanced Fv/Fm and chlorophyll content. The DAB assays suggested that these resistant phenotypes were also ROS-independent (H2O2 specifically) due to the strong positive impact of direct inhibition. The resistant phenotype and optimal efficacy of the aminochitosan MW fractions was observed at 3.5-15 kDa for antifungal efficacy and 15-20 kDa for in planta efficacy. Consequently, leaf senescence, hypersensitive responses and therefore necrosis were mitigated suggesting that aminochitosan primed defence responses in both mock and B. cinerea inoculated leaves. A temporal, label-free quantitative proteomic analysis revealed the differential priming of key molecular mechanisms underpinning aminochitosan primed states both with and without B. cinerea infection at 6 and 9 hpi in the tomato/B. cinerea pathosystem. Aminochitosan treatment (1 mg/mL) differentially regulated proteins as early as 6 hpi with some of the induced responses being sustained up to 9 hpi. Additionally, several proteins were oppositely regulated between aminochitosan pre-treatment and B. cinerea infection, indicating differential regulation patterns between the “primed state” and the “triggered state”. The proteomic data therefore partially validated the ‘priming' capacity of aminochitosan in 5- week-old tomato leaves, specifically diamino 3 when used as a foliar spray at a concentration of 1 mg/mL. The bimodal effects of aminochitosan in a maize/Fusarium verticillioides pathosystem were investigated by assessing the direct antifungal efficacy and elicitation of plant defence properties in planta. Aminochitosan displayed significant antifungal activity on both radial growth and sporulation at a minimum concentration of 1 mg/mL. Aminochitosan was also assessed in planta as preventative or curative treatments in maize seedlings infected with F. verticillioides at two time points. In the preventative treatment, salicylic acid accumulated during the early stages of infection (biotrophic phase) whereas in the curative treatment, jasmonic acid accumulated during the necrotrophic phase. In summary, we demonstrated that water-soluble aminochitosan possesses key properties that enable plant priming in addition to its superior direct antifungal activity compared to native chitosan. We also identified the optimal molecular weights and concentrations necessary for achieving maximum and 50% inhibition activity 2025-03-11T11:51:15Z 2025-03-11T11:51:15Z 2024 2025-03-11T11:49:05Z Thesis / Dissertation Doctoral PhD http://hdl.handle.net/11427/41146 Eng application/pdf Department of Molecular and Cell Biology Faculty of Science
spellingShingle Molecular and Cell Biology
Moola, Naadirah
6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
thesis_degree_str Doctoral
title 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
title_full 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
title_fullStr 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
title_full_unstemmed 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
title_short 6-deoxy-6-amino chitosan: A plant defence priming biopolymer that enhances resistance against Botrytis cinerea in tomato and Fusarium verticillioides in maize
title_sort 6 deoxy 6 amino chitosan a plant defence priming biopolymer that enhances resistance against botrytis cinerea in tomato and fusarium verticillioides in maize
topic Molecular and Cell Biology
url http://hdl.handle.net/11427/41146
work_keys_str_mv AT moolanaadirah 6deoxy6aminochitosanaplantdefenceprimingbiopolymerthatenhancesresistanceagainstbotrytiscinereaintomatoandfusariumverticillioidesinmaize