Development of a molecular biology tool kit to monitor dredging-related light stress in the seagrass Zostera muelleri ssp. capricorni in Port Curtis
Other Publication ResearchOnline@JCUAbstract
Key outcomes: This report provides a summary of field and laboratory studies conducted in the course of the development and validation of a set of molecular markers to monitor low light stress in the intertidal seagrass species Zostera muellerissp. capricorni. This is the first molecular study to establish and validate stress markers in an Australian seagrass species. The report also details how these markers can be used as a rapid assessment tool of seagrass health and be applied in active management of activities that impact on the light environment such as dredging. The key outcomes of these studies are: •We have successfully developed a seagrass health molecular tool kit based on real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) technology. This molecular tool kit was established by screening a total of 28 genes and includes a set of 10 reference genes and seven target genes (four early and three late sublethal markers of low light stress). •The molecular tool kit includes a combination of two types of gene responses to low light (up-regulated or down-regulated), which allows fast and robust diagnostics. We confirm plant gene expression returns to control levels once exposure to low light stress is removed. •The seagrass health molecular tool kit has been validated within a set of laboratory-based experiments and an intensive program of field collections, which includes shading studies as well as diel and monthly sampling at permanent transect sites within Port Curtis. •The newly developed field-to-bench workflow for analyzing seagrass health with this molecular tool kit, includes optimized seagrass sample handling in the field, processing, ribonucleic acid (RNA) extraction, cryostorage, bioinformatics and quality control through to the data analysis. The entire workflow has been developed for a fast turn-around time; typically within approx.1 working day upon receipt of field-collected samples. •The bioinformatics pipeline established at the University of Technology Sydney (UTS) to support this project is publicly available as a graphical-driven web-repository (http://144.6.226.79/), which will be used by scientists all over the world to assist in the examination of Z. muelleri transcriptomes. •This novel seagrass health molecular tool kit presents a significant advance in management of dredging operations to protect seagrasses because it: Can detect changes in Z. muelleri within 2 weeks of the onset of low light stress, long before any significant changes occur in physiology / morphology (8weeks); Substantially improves the capacity for reactive management by providing sufficient time for mitigating actions to be taken prior to seagrass loss; Allows assessments of stress to be made anywhere as they are not reliant on an established seagrass monitoring site with a history of known seagrass condition; and Provides a means to easily assess compliance with management conditions and effectiveness of management thresholds
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15/08
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33
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TropWATER
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Cairns, QLD, Australia
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