Publication Date

2024

Document Type

Dissertation/Thesis

First Advisor

Swingley, Wesley D.

Degree Name

Ph.D. (Doctor of Philosophy)

Legacy Department

Department of Biological Sciences

Abstract

Acaryochloris strains can adapt into far red region of the visible spectrum by utilizing its unique chlorophyll called chlorophyll d (Chl d). The genetic machinery behind Chl d has not been discovered yet. Biosynthetic pathway for other chlorophylls like chlorophyll a, chlorophyll b and chlorophyll f found in cyanobacteria are however well known. This research aims to understand the biosynthetic pathway responsible for chlorophyll d production. Bioinformatics approaches like comparative genomics, machine learning, and weighted gene co-expression network analysis (WGCNA) were central in this study to identify candidate genes and regulatory mechanisms potentially involved in chlorophyll d synthesis.

An important result of this study involved the successful draft genome assembly of Synechococcus species KUAC 3056, expanding the available genomic resources of basal strain to Acaryochloris clade. Comparative analysis of this basal strain with Chl d- lacking Acaryochloris thomasi strain RCC 1774, both of which lack chlorophyll d and plasmids, suggested connections between plasmid acquisition and chlorophyll d biosynthesis which was previously pointed out by other studies. Additionally, identified oxygenase-like and thiol-associated proteins present promising candidates for future experimental validation of their roles in Chl d synthesis.

Conjoint triad and Evolutionary Sequence Model (ESM) based feature extraction were performed for the cyanobacterial proteins to train random forest and neural network-based models. Resulting models have high accuracy and were deployed in Acaryochloris marina MBIC11017 to obtain potential candidate proteins involved in Chl d synthesis. The features embeddings of chlorophyll binding proteins were further studied to gain insights into the role of individual amino acids and their combination frequency in interacting with chlorophyll pigment. These models lay the groundwork for future integration with AlphaFold structural data, aiming to refine the predictive framework and enhance our understanding of chlorophyll-binding domain containing proteins.

WGCNA approach provided distinct co-expression modules, with yellow and blue modules displaying relatively strong associations with Acaryochloris strains. Genes within these modules, including photosystem, biosynthetic and hypothetical, might provide a clue to potential regulatory mechanisms of far-red oxygenic photosynthesis. Future work will include broader transcriptomic analyses across various light conditions, further elucidating the adaptive responses in Acaryochloris.

This research altogether advances the existing approach to understand chlorophyll d biosynthesis. The study also provides list of candidate genes which can hopefully be further studied using gene-knockout based studies to validate their potential role in Chl d synthesis.

Extent

210 pages

Language

en

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

Included in

Microbiology Commons

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