PYL1- and PYL8-like ABA Receptors of Nicotiana benthamiana Play a Key Role in ABA Response in Seed and Vegetative Tissue.

Gaston A Pizzio,Kacper P Kaminski,Cristian Mayordomo,Victor Garcia-Carpintero,Pedro L Rodriguez,Jorge Lozano-Juste,Maria Rivera-Moreno,Juan C Estevez,Nikolai V Ivanov,Marta Vazquez-Vilar,Diego Orzaez,Sergio G Nebauer,Armando Albert

doi:10.3390/cells11050795

Abstract

To face the challenges of climate change and sustainable food production, it is essential to develop crop genome editing techniques to pinpoint key genes involved in abiotic stress signaling. The identification of those prevailing abscisic acid (ABA) receptors that mediate plant-environment interactions is quite challenging in polyploid plants because of the high number of genes in the PYR/PYL/RCAR ABA receptor family. Nicotiana benthamiana is a biotechnological crop amenable to genome editing, and given the importance of ABA signaling in coping with drought stress, we initiated the analysis of its 23-member family of ABA receptors through multiplex CRISPR/Cas9-mediated editing. We generated several high-order mutants impaired in NbPYL1-like and NbPYL8-like receptors, which showed certain insensitivity to ABA for inhibition of seedling establishment, growth, and development of shoot and lateral roots as well as reduced sensitivity to the PYL1-agonist cyanabactin (CB). However, in these high-order mutants, regulation of transpiration was not affected and was responsive to ABA treatment. This reveals a robust and redundant control of transpiration in this allotetraploid plant that probably reflects its origin from the extreme habitat of central Australia.

Highlights

Nicotiana benthamiana (Nb) is a tobacco species widely used both in research and as a biotechnological crop for high-scale production of different bioproducts
To identify abscisic acid (ABA) receptors in the genome of Nb, we performed a BLAST search using as queries the amino acid sequences of the 14 Arabidopsis thaliana (At) PYR/PYL/REGULATORY COMPONENTS OF ABA RECEPTORS (RCAR) ABA receptors
As expected from previous analyses performed in At and different crops [10,11,23,24], we identified three major subfamilies of Nb ABA receptors, which we named as PYL1/2-like, PYL4/5-like, and PYL8/9-like

Summary

Introduction

Nicotiana benthamiana (Nb) is a tobacco species widely used both in research and as a biotechnological crop for high-scale production of different bioproducts. The Nicotiana genus is a part of Solanaceae, a family that includes economically relevant plants such as tomato (Solanum lycopersicum), potato (Solanum tuberosum), eggplant (Solanum melongena), and common tobacco (Nicotiana tabacum) [3]. The use of Agrobacterium-based transfection in Nb has been widely adopted for producing recombinant proteins (e.g., vaccines and antigens) as well as different biopharmaceuticals [4–6]. Nb has been established as an efficient biotechnological crop and a suitable model plant for studies on agronomic performance [6]. Given the increasing biotechnological relevance of Nb, several initiatives have been undertaken to develop genome sequencing and transcriptome analysis of Nb [7,8]. Nb is an allotetraploid tobacco whose diploid parental progenitors (hybridization event 4–5 million years ago [Mya]) are affiliated with the Nicotiana sections

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Conclusion