Chloroplasts are attractive platforms for synthetic biology applications since they are capable of driving very high levels of transgene expression, if mRNA production and stability are properly regulated. However, plastid transformation is a slow process and currently limited to a few plant species. The liverwort Marchantia polymorpha is a simple model plant that allows rapid transformation studies; however, its potential for protein hyperexpression has not been fully exploited. This is partially due to the fact that chloroplast post-transcriptional regulation is poorly characterized in this plant. We have mapped patterns of transcription in Marchantia chloroplasts. Furthermore, we have obtained and compared sequences from 51 bryophyte species and identified putative sites for pentatricopeptide repeat protein binding that are thought to play important roles in mRNA stabilization. Candidate binding sites were tested for their ability to confer high levels of reporter gene expression in Marchantia chloroplasts, and levels of protein production and effects on growth were measured in homoplastic transformed plants. We have produced novel DNA tools for protein hyperexpression in this facile plant system that is a test-bed for chloroplast engineering.

The hornworts are a small group of land plants, consisting of only 11 families and approximately 220 species. Despite their small size as a group, their phylogenetic position and unique biology are of great importance. Hornworts, together with mosses and liverworts, form the monophyletic group of bryophytes that is sister to all other land plants (Tracheophytes). It is only recently that hornworts became amenable to experimental investigation with the establishment of Anthoceros agrestis as a model system. In this perspective, we summarize the recent advances in the development of A. agrestis as an experimental system and compare it with other plant model systems. We also discuss how A. agrestis can help to further research in comparative developmental studies across land plants and to solve key questions of plant biology associated with the colonization of the terrestrial environment. Finally, we explore the significance of A. agrestis in crop improvement and synthetic biology applications in general.

The rising population and the ongoing climate crisis call for improved means to monitor and optimise agriculture. A promising approach to tackle current challenges in food production is the early diagnosis of plant diseases through non-invasive methods, such as the detection of volatiles. However, current devices for detection of multiple volatiles are based on electronic noses, which are expensive, require complex circuit assembly, may involve metal oxides with heating elements, and cannot easily be adapted for some applications that require miniaturisation or limit front-end use of electronic components. To address these challenges, a low-cost optoelectronic nose using chemo-responsive colorimetric dyes drop-casted onto filter paper has been developed in the current work. The final sensors could be used for the quantitative detection of up to six plant volatiles through changes in colour intensities with a sub-ppm level limit of detection, one of the lowest limits of detection reported so far using colorimetric gas sensors. Sensor colouration could be analysed using a low-cost spectrometer and the results could be processed using a microcontroller. The measured volatiles could be used for the early detection of plant abiotic stress as early as two days after exposure to two different stresses: high salinity and starvation. This approach allowed a lowering of costs to GBP 1 per diagnostic sensing paper. Furthermore, the small size of the paper sensors allows for their use in confined settings, such as Petri dishes. This detection of abiotic stress could be easily achieved by exposing the devices to living plants for 1 h. This technology has the potential to be used for monitoring of plant development in field applications, early recognition of stress, implementation of preventative measures, and mitigation of harvest losses.

Chloroplast genes are present at high ploidy in plants, and capable of driving very high levels of gene expression if mRNA production and stability are properly regulated. Marchantia polymorpha is a simple model plant that allows rapid transformation studies, however post-transcriptional regulation in plastids is poorly characterized in this liverwort. We have mapped patterns of transcription in Marchantia chloroplasts. Furthermore, we have obtained and compared sequences from 51 early-divergent plant species, and identified putative sites for pentatricopeptide repeat protein binding that are thought to play important roles in mRNA stabilisation. Candidate binding sites were tested for their ability to confer high levels of reporter gene expression in Marchantia chloroplasts, and levels of protein production and effects on growth were measured in homoplasmic transformed plants. We have produced novel DNA tools for protein hyper-expression in a facile plant system that is a test-bed for chloroplast engineering.