Transient expression in Nicotiana benthamiana for rapid functional analysis of genes involved in non-photochemical quenching and carotenoid biosynthesis
| Autor: | Leonelli, Lauriebeth, Erickson, Erika, Lyska, Dagmar, Niyogi, Krishna K |
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| Rok vydání: | 2016 |
| Předmět: |
photosynthesis
fungi Plant Biology & Botany food and beverages Thalassiosira transient assay xanthophyll cycle Plant Biology Original Articles PSBS Xanthophylls non‐photochemical quenching Carotenoids lutein epoxide Affordable and Clean Energy Tobacco Genetics Nicotiana benthamiana Original Article Nannochloropsis Biochemistry and Cell Biology carotenoid biosynthesis non-photochemical quenching Biotechnology Plant Proteins |
| Zdroj: | The Plant journal : for cell and molecular biology, vol 88, iss 3 The Plant Journal |
| Popis: | Summary Plants must switch rapidly between light harvesting and photoprotection in response to environmental fluctuations in light intensity. This switch can lead to losses in absorbed energy usage, as photoprotective energy dissipation mechanisms can take minutes to hours to fully relax. One possible way to improve photosynthesis is to engineer these energy dissipation mechanisms (measured as non‐photochemical quenching of chlorophyll a fluorescence, NPQ) to induce and relax more quickly, resulting in smaller losses under dynamic light conditions. Previous studies aimed at understanding the enzymes involved in the regulation of NPQ have relied primarily on labor‐intensive and time‐consuming generation of stable transgenic lines and mutant populations – approaches limited to organisms amenable to genetic manipulation and mapping. To enable rapid functional testing of NPQ‐related genes from diverse organisms, we performed Agrobacterium tumefaciens‐mediated transient expression assays in Nicotiana benthamiana to test if NPQ kinetics could be modified in fully expanded leaves. By expressing Arabidopsis thaliana genes known to be involved in NPQ, we confirmed the viability of this method for studying dynamic photosynthetic processes. Subsequently, we used naturally occurring variation in photosystem II subunit S, a modulator of NPQ in plants, to explore how differences in amino acid sequence affect NPQ capacity and kinetics. Finally, we functionally characterized four predicted carotenoid biosynthesis genes from the marine algae Nannochloropsis oceanica and Thalassiosira pseudonana and examined the effect of their expression on NPQ in N. benthamiana. This method offers a powerful alternative to traditional gene characterization methods by providing a fast and easy platform for assessing gene function in planta. Significance Statement Transient expression of genes in Nicotiana benthamiana has become a method of choice to quickly assess gene function in planta. Here we illustrate the potential of transient assays in N. benthamiana to provide a medium‐throughput platform for assessing gene function with regards to photosynthetic processes. A promising system is described for studying the regulation of NPQ dynamics toward minimizing energy losses and improving photosynthetic yields. |
| Databáze: | OpenAIRE |
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