A high-throughput transient expression system for rice

Page, Michael and Parry, Martin Afan John and Carmo-Silva, Ana Elizabete (2019) A high-throughput transient expression system for rice. Plant, Cell and Environment, 42 (7). pp. 2057-2064. ISSN 0140-7791

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Abstract

Rice is an important global crop and represents a vital source of calories for many food insecure regions. Efforts to improve this crop by improving yield, nutritional content, stress tolerance, or resilience to climate change are certain to include biotechnological approaches, which rely on the expression of transgenes in planta. The throughput and cost of currently available transgenic expression systems is frequently incompatible with modern, high-throughput molecular cloning methods. Here, we present a protocol for isolating high yields of green rice protoplasts and for PEG-mediated transformation of isolated protoplasts. Factors affecting transformation efficiency were investigated, and the resulting protocol is fast, cheap, robust, high-throughput, and does not require specialist equipment. When coupled to a high-throughput modular cloning system such as Golden Gate, this transient expression system provides a valuable resource to help break the "design-build-test" bottleneck by permitting the rapid screening of large numbers of transgenic expression cassettes prior to stable plant transformation. We used this system to rapidly assess the expression level, subcellular localisation, and protein aggregation pattern of nine single-gene expression cassettes, which represent the essential component parts of the beta-cyanobacterial carboxysome.

Item Type:
Journal Article
Journal or Publication Title:
Plant, Cell and Environment
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/1300/1314
Subjects:
ID Code:
131793
Deposited By:
Deposited On:
06 Mar 2019 09:30
Refereed?:
Yes
Published?:
Published
Last Modified:
24 Nov 2020 07:16