Rational design of minimal synthetic promoters for plants

Cai, Yuyang; Kallam, Kalyani; Tidd, Henry; Gendarini, Giovanni; Salzman, Amanda; Patron, Nicola J.

doi:10.1093/nar/gkaa682

Cited by 88 publications

(98 citation statements)

References 72 publications

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“…Additionally, there are already examples of short, strong constitutive promoters from Arabidopsis, such as AtTCTP (0.3 bp ( 77)) and AtSCPL30 (0.45bp ( 78)). A recent study was also introduced short synthetic promoters for plant expression (79).…”

Section: New Libraries Of Short Promoter and Terminator Partsmentioning

confidence: 99%

Mobius Assembly for Plant Systems highlights promoter-terminator interaction in gene regulation

Andreou

Nirkko

Ochoa-Villarreal

et al. 2021

Preprint

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Plant synthetic biology is a fast-evolving field that employs engineering principles to empower research and bioproduction in plant systems. Nevertheless, in the whole synthetic biology landscape, plant systems lag compared to microbial and mammalian systems. When it comes to multigene delivery to plants, the predictability of the outcome is decreased since it depends on three different chassis: E.coli, Agrobacterium, and the plant species. Here we aimed to develop standardised and streamlined tools for genetic engineering in plant synthetic biology. We have devised Mobius Assembly for Plant Systems (MAPS), a user-friendly Golden Gate Assembly system for fast and easy generation of complex DNA constructs. MAPS is based on a new group of small plant binary vectors (pMAPs) that contains an origin of replication from a cryptic plasmid of Paracoccuspantotrophus. The functionality of the pMAP vectors was confirmed by transforming the MM1 cell culture, demonstrating for the first time that plant transformation is dependent on the Agrobacterium strains and plasmids; plasmid stability was highly dependent on the plasmid and bacterial strain. We made a library of new short promoters and terminators and characterised them using a high-throughput protoplast expression assay. Our results underscored the strong influence of terminators in gene expression, and they altered the strength of promoters in some combinations and indicated the presence of synergistic interactions between promoters and terminators. Overall this work will further facilitate plant synthetic biology and contribute to improving its predictability, which is challenged by combinatorial interactions among the genetic parts, vectors, and chassis.

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Section: New Libraries Of Short Promoter and Terminator Partsmentioning

confidence: 99%

Mobius Assembly for Plant Systems highlights promoter-terminator interaction in gene regulation

Andreou

Nirkko

Ochoa-Villarreal

et al. 2021

Preprint

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“…A strategy to alleviate deleterious growth effects would consist in disconnecting plant growth from pheromone production. This could be done by employing agronomically-compatible inducible expression systems for the activation of the pathway, taking advantage of the increasingly number of Synthetic Biology tools made available for plants and particularly for Nicotiana species (Bernabé-Orts et al, 2020;Cai et al, 2020;Molina-Hidalgo et al, 2020). Alternatively, the use of a different plant chassis displaying specialized structures such as glandular trichomes to store potentially toxic pheromone compounds could be advantageous.…”

Section: Discussionmentioning

confidence: 99%

Production of Volatile Moth Sex Pheromones in TransgenicNicotiana benthamianaPlants

Mateos-Fernández

Moreno-Giménez

Gianoglio

et al. 2021

Preprint

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Plant-based bio-production of insect sex pheromones has been proposed as an innovative strategy to increase the sustainability of pest control in agriculture. Here we describe the engineering of transgenic plants producing (Z)-11-hexadecen-1-ol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multiple multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them in Nicotiana benthamiana plants. The constructs comprised the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera farnesyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, whose severity correlated with pheromone levels. All but one of the recovered lines produced high levels of Z11-16OH but very low levels of Z11-16OAc, probably as a result of recurrent truncations at the level of the EaDAct gene. Only one plant line (SxPv1.2) was recovered harbouring an intact pheromone pathway and producing moderate levels of Z11-16OAc (11.8 μg g-1 FW), next to high levels of Z11-16OH (111.4 μg g-1). Z11-16OAc production was accompanied in SxPv1.2 by a partial recovery of the dwarf phenotype. SxPv1.2 was used to estimate the rates of volatile pheromone release, which resulted in 8.48 ng g-1 FW per day for Z11-16OH and 9.44 ng g-1 FW per day for Z11-16OAc. Our results suggest that pheromone release acts as a limiting factor in pheromone bio-dispenser strategies and establish a roadmap for biotechnological improvements.

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“…Expression under strong viral constitutive promoter CaMV 35S can experience silencing or patterns in dividing cells [ 7 ] and is therefore often exchanged with plant constitutive promoters, such as rice actin and maize ubiquitin promoters. Still, variable expression in different species forces the exploration of novel options [ 154 , 155 ]. On the other hand, it is possible to opt for separation of two or more reporters at the protein stage.…”

Section: Challenges Of Plant Biosensors’ Development and Usementioning

confidence: 99%

Biosensors: A Sneak Peek into Plant Cell’s Immunity

Levak

Lukan

Gruden

et al. 2021

Life

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Biosensors are indispensable tools to understand a plant’s immunity as its spatiotemporal dimension is key in withstanding complex plant immune signaling. The diversity of genetically encoded biosensors in plants is expanding, covering new analytes with ever higher sensitivity and robustness, but their assortment is limited in some respects, such as their use in following biotic stress response, employing more than one biosensor in the same chassis, and their implementation into crops. In this review, we focused on the available biosensors that encompass these aspects. We show that in vivo imaging of calcium and reactive oxygen species is satisfactorily covered with the available genetically encoded biosensors, while on the other hand they are still underrepresented when it comes to imaging of the main three hormonal players in the immune response: salicylic acid, ethylene and jasmonic acid. Following more than one analyte in the same chassis, upon one or more conditions, has so far been possible by using the most advanced genetically encoded biosensors in plants which allow the monitoring of calcium and the two main hormonal pathways involved in plant development, auxin and cytokinin. These kinds of biosensor are also the most evolved in crops. In the last section, we examine the challenges in the use of biosensors and demonstrate some strategies to overcome them.

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Rational design of minimal synthetic promoters for plants

Cited by 88 publications

References 72 publications

Mobius Assembly for Plant Systems highlights promoter-terminator interaction in gene regulation

Mobius Assembly for Plant Systems highlights promoter-terminator interaction in gene regulation

Production of Volatile Moth Sex Pheromones in TransgenicNicotiana benthamianaPlants

Biosensors: A Sneak Peek into Plant Cell’s Immunity

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