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Sexual Plant Reproduction – Seed formation

Periodic Reporting for period 2 - SexSeed (Sexual Plant Reproduction – Seed formation)

Reporting period: 2018-03-01 to 2020-02-29

To feed a growing world population, it will be necessary to increase agricultural yields, without increasing the amount of arable land.
Understanding the factors that regulate sexual reproduction will enable this critical aspect of agricultural production to be engineered. Our project will provide new insights into the network controlled by SEEDSTICK (STK), a MADS box transcription factor, which is the master regulator of seed production. The objective of this proposal was to strengthen research partnerships through staff exchanges and networking activities, at international and intercontinental levels. By combining transcriptomic and genetic approaches, we aimed to uncover new functions for STK targets and implicate them in putative signalling cascades, increasing our knowledge on the network that controls seed formation in Arabidopsis. Arabidopsis is an excellent model to study seed formation, as it shares a conserved developmental program with major seed-producing crop plants, important to improve not only gross agricultural productivity, but also the composition of seeds and hence the production of components used for high added-value seed-derived products. The growing importance of seeds and seed-derived products to humanity and the central role of STK in seed development means that this project has great potential to contribute to Europe’s excellence and competitiveness in the world.
The objective was to better understand the factors that regulate sexual reproduction in plants and enable increased productivity through engineered agricultural production. Very high quality results were obtained. WP2 (comparison of the pattern of expression of the STK gene by RNAseq to ChIPseq data) was achieved and following WPs started according to the work plan. Analysis of results was performed. Additionally, this new data was confronted with RNA-sequencing data from a stk mutant (Mizzotti et al., 2014) allowing the differentiation of direct targets that were also differentially expressed (up or down-regulated). The validation of the targets was accessed by qRT-PCR.
Functional analyses of selected genes was done through the study of several mutants regarding different reproductive stages: 1) early phases of ovule development, 2) fertilization process, 3) seed/fruit development. For the targets with no mutants available in the public databases, RNA interference and CRISPR/Cas9 was produced as an alternative approach.
The objectives initially proposed were achieved and exceeded in some of the tasks, such as WP3, Functional Characterization of the selected target genes involved in early stages of ovule development and WP4, Functional Characterization of the selected target genes related to fertilization. The results obtained with the project are of high scientific quality as can be evaluated by the scientific production, with 47 presentations at national and International Conferences and workshops, 12 Publications in Scientific Journals and more than 12 still in preparation.
The scientific impact was very good quality. The impact on the training and career perspectives of the young researchers was excellent. Young European researchers were trained in non- EU highly specialised laboratories and they are bringing this knowledge back to Europe. The long-term impact is still too early to fully evaluate but very promising. Thanks to the networking between participants, researchers have been able to finalise several masters and PhD- theses. One researcher received a MSCA Fellowship to be implemented within the consortium and another researcher received a job offer in the industry. Young researchers have all stressed how good the secondments were for their training.
The project contributed to the development of lasting collaborations between top international groups within and outside Europe in the field of engineered agricultural production thus reinforcing the position of the EU on such topic. This will be translated into top scientific publications and attract the interest of the general public.
Competitiveness of the researchers had clearly improved as they had access to state-of-the-art facilities, protocols and materials that are also greatly enhancing their training capacities.
Common publications between the different members of the consortium are now being submitted. 4 workshops (Tucson, Padova, Porto and Adelaide) and 1 congress (Padova) took place and 2 satellite meetings were prepared for the summer 2018 (Florence and Nagoya).
The website was continuously updated and activities reported for a broader audience (Fascinating Day Milano, IJUP Porto, Summer School for young students (between 8 - 18 y.o.) acknowledging EU funding.
Most agricultural production is aimed at the production of seeds through sexual reproduction, and these seeds account for the majority of the calories consumed by humans. The project when completed will allow increasing seed production in highly nutritive plants for food and feed. Therefore, it addresses major societal- and worldwide development- needs. It is expected in the future, to enhance innovation capacity through technological development and new products and create market opportunities while strengthening competitiveness and growth of companies in a changing climate and environment.

The benefits of the project financed throughout have been not only important for the research groups and institutions, but mainly for the students at the European institutions. The different ERS that participate in this international consortium, profited from internationalization which would not be possible without the project.
In the last four years, all post-graduate students in the partner laboratories had the opportunity to intern in the best laboratories in the field of reproductive biology in the United States, Japan and Australia, with grants funded by their European project SexSeed for periods of 2 to 12 months. During these periods students acquired a set of technical skills; bioinformatics, molecular biology, genetic engineering, bio-imaging, etc., impossible to achieve in an equivalent way in their laboratories, with the added value of being able to transfer all this know-how to their research laboratory, institution and other students involved. In the SexSeed project, the research work developed focuses on the study of the molecular network controlled by SEEDSTICK (STK), a MADS type transcription factor, the main regulator of seed development. The project essentially aimed to strengthen international research partnerships through the exchange of students and researchers and networking activities, at international and intercontinental level.
The achievements are well measurable by the Exploitation and Dissemination deliverables presented in the portal and by the already achieved results and the ones still underway, just presented.
pAGP31::GUS
pKNU::YFP