Research Article

Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field

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Science  04 Jan 2019:
Vol. 363, Issue 6422, eaat9077
DOI: 10.1126/science.aat9077

Fixing photosynthetic inefficiencies

In some of our most useful crops (such as rice and wheat), photosynthesis produces toxic by-products that reduce its efficiency. Photorespiration deals with these by-products, converting them into metabolically useful components, but at the cost of energy lost. South et al. constructed a metabolic pathway in transgenic tobacco plants that more efficiently recaptures the unproductive by-products of photosynthesis with less energy lost (see the Perspective by Eisenhut and Weber). In field trials, these transgenic tobacco plants were ∼40% more productive than wild-type tobacco plants.

Science, this issue p. eaat9077; see also p. 32

Structured Abstract

INTRODUCTION

Meeting food demands for the growing global human population requires improving crop productivity, and large gains are possible through enhancing photosynthetic efficiency. Photosynthesis requires the carboxylation of ribulose-1,5-bisphosphate (RuBP) by ribulose-1,5-bisphosphate carboxylase-oxygenase (RuBisCO), but photorespiration occurs in most plants such as soybean, rice, and wheat (known as C3 crops) when RuBisCO oxygenates RuBP instead, requiring costly processing of toxic byproducts such as glycolate. Photorespiration can reduce C3 crop photosynthetic efficiency by 20 to 50%. Although various strategies exist for lowering the costs of photorespiration, chamber- and greenhouse-grown plants with altered photorespiratory pathways within the chloroplast have shown promising results, including increased photosynthetic rates and plant size.

RATIONALE

To determine if alternative photorespiratory pathways could effectively improve C3 field crop productivity, we tested the performance of three alternative photorespiratory pathways in field-grown tobacco. One pathway used five genes from the Escherichia coli glycolate oxidation pathway; a second pathway used glycolate oxidase and malate synthase from plants and catalase from E. coli; and the third pathway used plant malate synthase and a green algal glycolate dehydrogenase. All enzymes in the alternative pathway designs were directed to the chloroplast. RNA interference (RNAi) was also used to down-regulate a native chloroplast glycolate transporter in the photorespiratory pathway, thereby limiting metabolite flux through the native pathway. The three pathways were introduced with and without the transporter RNAi construct into tobacco, which is an ideal model field crop because it is easily transformed, has a short life cycle, produces large quantities of seed, and develops a robust canopy similar to that of other field crops.

RESULTS

Using a synthetic biology approach to vary promoter gene combinations, we generated a total of 17 construct designs of the three pathways with and without the transporter RNAi construct. Initial screens for photoprotection by alternative pathway function under high–photorespiratory stress conditions identified three to five independent transformants of each design for further analysis. Gene and protein expression analyses confirmed expression of the introduced genes and suppression of the native transporter in RNAi plants. In greenhouse screens, pathway 1 increased biomass by nearly 13%. Pathway 2 showed no benefit compared to wild type. Introduction of pathway 3 increased biomass by 18% without RNAi and 24% with RNAi, which were consistent with changes in photorespiratory metabolism and higher photosynthetic rates. Ultimately, field testing across two different growing seasons showed significant increases in biomass of pathway 3 plants with RNAi compared to WT of 20% in 2016 (P =0.04) and by 24% in 2017 (P = 0.018). In addition, this pathway increased the light-use efficiency of photosynthesis by 17% in the field.

CONCLUSION

Engineering more efficient photorespiratory pathways into tobacco while inhibiting the native pathway markedly increased both photosynthetic efficiency and vegetative biomass. We are optimistic that similar gains may be achieved and translated into increased yield in C3 grain crops because photorespiration is common to all C3 plants and higher photosynthetic rates under elevated CO2, which suppresses photorespiration and increases harvestable yield in C3 crops.

Alternative photorespiratory pathways in tobacco.

Three alternative pathways [1 (red), 2 (dark blue), and 3(light blue)] introduced into tobacco chloroplasts for more efficient recycling of glycolate. RNAi suppresses the native glycolate/glycerate transporter PLGG1 to prevent glycolate from leaving the chloroplast and entering the native pathway (gray).

Abstract

Photorespiration is required in C3 plants to metabolize toxic glycolate formed when ribulose-1,5-bisphosphate carboxylase-oxygenase oxygenates rather than carboxylates ribulose-1,5-bisphosphate. Depending on growing temperatures, photorespiration can reduce yields by 20 to 50% in C3 crops. Inspired by earlier work, we installed into tobacco chloroplasts synthetic glycolate metabolic pathways that are thought to be more efficient than the native pathway. Flux through the synthetic pathways was maximized by inhibiting glycolate export from the chloroplast. The synthetic pathways tested improved photosynthetic quantum yield by 20%. Numerous homozygous transgenic lines increased biomass productivity between 19 and 37% in replicated field trials. These results show that engineering alternative glycolate metabolic pathways into crop chloroplasts while inhibiting glycolate export into the native pathway can drive increases in C3 crop yield under agricultural field conditions.

This is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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