Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - Newcotiana (Developing Multipurpose Nicotiana Crops for Molecular Farming using New Plant Breeding Techniques)

Teaser

NEWCOTIANA is a Research project that employs New Breeding Techniques to develop new plant varieties of the genus Nicotiana producing end-value chemicals including proteins and metabolites. Plants can be used as biofactories of added-value chemicals, e.g. biopharmaceuticals...

Summary

NEWCOTIANA is a Research project that employs New Breeding Techniques to develop new plant varieties of the genus Nicotiana producing end-value chemicals including proteins and metabolites. Plants can be used as biofactories of added-value chemicals, e.g. biopharmaceuticals, at an agricultural scale, offering a sustainable solution for present and future manufacturing needs. The solanaceous plant species within the genus Nicotiana, more specifically Nicotiana tabacum (cultivated tobacco) and Nicotiana benthamiana (an Australian tobacco relative), are among the most widely used plant biofactories because of their favorable attributes, which include high metabolic versatility, ease of cultivation and high yield, availability of genetic tools for trait manipulation, and their non-food status, which minimizes the possibility of contamination of the food supply with industrial designated products.

In stark contrast with its high appreciation as biofactories, traditional tobacco cultivation is in continuous decline in Europe due, among other reasons, to the bad reputation of a crop that is used mainly for manufacturing cigarettes and other smoking products. The aim of NEWCOTIANA project is to offer an alternative to tobacco farmers by breeding competitive Nicotiana biofactory varieties for high value non-smoking products. This would create a suitable production platform for the EU bioeconomy and at the same time would bring sustainability to a traditional crop. However, to become fully competitive and scalable biofactories, Nicotiana plants need to incorporate unconventional traits such as protein stability or metabolite fortification, which are not accessible to traditional crop breeding. Recently, New Plant Breeding Techniques (NPBTs) have emerged that bring unprecedented opportunities for Plant Biotechnology. The driving hypothesis in the NEWCOTIANA project is that breeding tobacco biofactory varieties for high value non-smoking products is now possible with the use of NPBTs. Four of those NPBTs, namely sequence-directed nuclease (SDN)-mediated genome editing (popularly known as CRISPR), cis/intra-genesis, agroinfiltration and grafting, will be employed in the project.

Most importantly, NEWCOTIANA goals include the improvement of the awareness and public understanding of New Plant Breeding Technologies. Our aim is to create a two-way dialogue between science on the one hand and stakeholders, policymakers and the general public on the other, through provision of unbiased information and reliable data arising from the project, through innovative and highly participative channels. In this way, NEWCOTIANA will provide industry, policy-makers and consumers with experimental evidence and social communication channels to facilitate the decision-making process for the adoption of NPBTs.

Work performed

During the course of the first 18 months of the project a number of important results have been already achieved:

- A top quality Nicotiana benthamiana genome assembly has been accomplished, with chromosome-level assembly scaffolds and Webtool Apollo integration that allows community annotation on the genes. This genome assembly will allow researchers to better design their breeding strategies, select CRISPR targets and will provide important clues on the genetic basis of N. benthamiana favorable biofactory traits.

- An extended toolbox for NPBTs in N. benthamiana has been created, incorporating new tools for multiplexing editing, newly adapted CRISPR enzymes, advances in Gene Transfer, and new intragenic markers for Nicotiana.

- Significant advances have been made in the generation of new N. benthamiana and N. tabacum biofactory lines. As planned for this period, a number of genetic factors influencing recombinant protein quality have been identified. In parallel, the generation of new plants with favorable metabolic traits is progressing adequately. Plants lines with multiple CRISPRed genes (up to 7 knock-out genes in a single plant generation!), have been generated, and species are being generated using horizontal genome transfer (grafting) technology.

- Active communication/dissemination campaign: Initial press release reached high impact; more than 75 communication and dissemination activities have been performed including videos, conferences, open workshops, public exhibitions, training schools. Brand identity guidelines in webpage and Social media (Twitter 317 followers) were established; Seven research articles published.

Final results

At this stage NEWCOTIANA has already advanced beyond the state-of-the-art in sequence-specific nuclease (SSN) technologies and platform optimization. Regarding SSNs technologies, we have developed alternative nucleases and advanced in faster delivery systems and more efficient gene editing/ targeting methods. Regarding platform optimization, the new N. benthamiana genome assembly has set a crucial milestone in plant biofactories, moving forward the field to a new level that will allow creation of prototypes incorporating new valuable traits on ER-stability, protease activity, glycosylation, RNA silencing, biomass composition or biocontainment. In addition to the current advances, the following impacts are expected from the project.

Scientific impact: the project will deepen in the understanding of gene function in Nicotiana and the interactions between the metabolome and agronomic behavior. NEWCOTIANA will produce improved NPBT tools for plant breeding that are more efficient and accurate than those currently available.

Technical impact: NEWCOTIANA will create a number of elite plant varieties (demonstrators) that will serve as performance indicators. A demonstrator should produce functional recombinant protein with higher quality and reduced losses due to proteolytic degradation than current N. benthamiana platforms. Other demonstrator should consist in a N. tabacum plant producing competitive quantities of added value metabolites such as e.g. anatabine, zeaxanthin or squalene in seeds.

Social impact: In the EU, there are approximately 60.000 tobacco farmers and as many indirect jobs threatened by the decline in tobacco demand. NEWCOTIANA will bring the opportunity to maintain those jobs and to generate new ones, revitalizing rural areas where tobacco is a traditional crop as well as new others that will incorporate NEWCOTIANA. In a wider perspective, the scalability of the NEWCOTIANA biofactory will improve accessibility to biopharmaceuticals (vaccines, monoclonal antibodies and others) whose availability is endangered due to high manufacturing costs and/or long production cycles. Furthermore, NEWCOTIANA aims to provide industry, policy-makers and consumers with the experimental evidence that will facilitate the decision-making process for the adoption of NPBTs.

Website & more info

More info: https://newcotiana.org/.