Title: Transient expression in Nicotiana benthamiana for rapid functional analysis of genes involved in non‐photochemical quenching and carotenoid biosynthesis

Abstract

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 acidmore » 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 .« less

Authors:

Leonelli, Lauriebeth ^[1]; Erickson, Erika ^[2]; Lyska, Dagmar ^[2]; Niyogi, Krishna K. ^[2]

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+ Show Author Affiliations

1. Howard Hughes Medical Institute Department of Plant and Microbial Biology University of California Berkeley CA 94720‐3102 USA
2. Howard Hughes Medical Institute Department of Plant and Microbial Biology University of California Berkeley CA 94720‐3102 USA, Molecular Biophysics and Integrated Bioimaging Division Lawrence Berkeley National Laboratory Berkeley CA 94720 USA

Publication Date:

Thu Sep 15 00:00:00 EDT 2016

Research Org.:

Univ. of California, Oakland, CA (United States)

Sponsoring Org.:

USDOE Advanced Research Projects Agency - Energy (ARPA-E)

OSTI Identifier:

1324846

Alternate Identifier(s):

OSTI ID: 1325380; OSTI ID: 1625938

Grant/Contract Number:  

DE‐AR0000204; AR0000204

Resource Type:

Published Article

Journal Name:

The Plant Journal

Additional Journal Information:

Journal Name: The Plant Journal Journal Volume: 88 Journal Issue: 3; Journal ID: ISSN 0960-7412

Publisher:

Wiley-Blackwell

Country of Publication:

United Kingdom

Language:

English

Subject:

Plant Sciences