A flower is an anatomically complex organ containing tissues of different identity which undergo distinct
developmental programs but which have to be synchronized to ensure successful gamete formation
and fertilization. Stress conditions have diverse effects on different tissues, but also the sensitivity for a
specific tissue or cell type can vary among the different developmental stages.
One of the major challenges in the field of stress resilience in reproductive tissues is the dissection of
the phenotype on the tissue and cellular level. This information is important for the characterization of
the basis of the sensitivity and the selection of specific targets (e.g. cell types or tissues) for
improvement. Due to the easier access to pollen, stress related studies have mainly focused on the
resilience of the male gametophyte, often ignoring the female counterpart or even the vegetative tissues
of the flower organ. As a consequence, a detailed picture of the effects of abiotic stresses both on the
response and the tolerance of the different flower tissues is absent.
WG1 will lead the efforts to unify, optimize and standardize protocols used for studying stress resilience
and response in reproductive tissues in various model and crop plants, propose novel methods for
phenotyping and create tools for –omics data acquisition and analysis. TASK 1.1: Determine the critical information required to decipher stress response and resilience
in reproductive tissues. State-of-the-art –omics and phenotypic techniques need to be utilised in order
to provide an in-depth description of the response of different male and female reproductive tissues to
different and combined stress scenarios. Only a basic set of such techniques has been used so far and
therefore the picture of stress response is far from being complete. Experts from different fields of stress
biology and different –omics strategies (including GWAS - genome-wide association studies) will
describe the missing levels of information and the steps that need to be taken to establish a
comprehensive description of the stress response. In addition, members of WG1 will define the
limitations in the current approaches for stress resilience determination in plant reproduction and will
propose novel techniques to address resilience under laboratory and field conditions, both for
reproductive success and yield. TASK 1.2: Optimize and standardize methods for monitoring stress response and resilience in
reproductive tissues in major crops. Currently different laboratories use different methods to define
stress tolerance in reproductive tissues. Members of WG1 will define a commonly accepted set of
methods that can be used to verify stress resilience in reproductive tissues and yield across different
levels of experimental scales, including laboratory and field trials. These methods will ensure
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reproducibility, and minimize man made error in data collection and analysis. Pitfalls and limitations of
these techniques will be identified and measures for their improvement will be determined. For each
crop, stress resilience thresholds will be defined under different conditions, which will allow a direct
comparison of results across different laboratories. By this, we aim to create a common experimental
“language” that will boost cooperation, cross-validation, and progress in the field. The improved
protocols will be benchmarked by different groups and data will be submitted for evaluation using a
RECROP platform based on a blind analysis approach. By this RECROP will develop pipelines for stress
application, data collection and analysis and result interpretation. The pipelines and the results from the
benchmarking will be reported in the meetings and conference organized by RECROP, and will be
published as a peer reviewed open access method manuscripts. TASK 1.3: Advanced imaging techniques to dissect the effects of abiotic stresses on
reproductive tissues at the cellular level. Improving resilience either by genetic approaches or
agricultural means requires a prior knowledge of the basis of sensitivity. Therefore, it is of utmost
importance to define on the tissue and cell-type levels the effects of different stress combinations, as
sensitive cells can be targeted for genetic improvement. RECROP will provide a framework of various
bioimaging techniques for the 3D analysis of stressed floral meristems. RECROP will include photon-
based tomography techniques such as confocal, light sheet and super-resolution microscopy, X-ray
microscopy (XRM) and Magnetic resonance imaging (MRI) to create 3D images of stressed flowers to
capture defects or alterations in specific tissues due to stress. Bioimages will be used to create models
for stress effects on reproductive tissues based on deep learning approaches that will provide robust
automated algorithms to develop a 3D Digital Tissue Atlas for different crops exposed to different
stresses. Such analysis will allow us to decipher the most sensitive cells and tissues during reproductive
development under different stress conditions. TASK 1.4: Create a common platform for integration and analysis of –omics datasets from abiotic
stress studies. WG1 will coordinate the integration of existing and upcoming –omics datasets in a
single platform accessible through the RECROP website. The platform will operate under a user-friendly
interface for biologists and non-experts and will provide comparative analysis of RNAs, proteins, and
metabolites between different conditions, tissues and species as well as multilevel analysis of specific
pathways. New data generated during the RECROP will be integrated into the platform to increase
statistical power. WG1 members will create a commonly accepted pipeline for data acquisition, quality
check, integration, and analysis. The platform will allow the easy mining of the results which will be
visualised as ready to use images for publications or presentations. The platform will be presented in
COST meetings, conferences and a publication, to encourage more researchers to deposit relevant
data. Students and COST-members will be trained on the use of the platform through workshops and
webinars. We envision that the platform will operate beyond the funding period of RECROP.
Milestones WG1: M1.1 (Y1M12) List with methods and layers of information required to characterize crop stress resilience; M1.2 (Y2M12) Agree on commonly accepted and optimized protocols; M1.3 (Y3M3) Publication of method article on bioimaging and 3D modelling of floral meristem tissues; M1.4 (Y3M3) Online and fully operating platform for integration of –omics datasets from different crops; M1.5 (Y4M2) benchmarking and open access publication of the platform on a scientific journal
WG1 Contact: Christos Bazakos – cbazakos@elgo.gr
Milestones WG1: M1.1 (Y1M12) List with methods and layers of information required to characterize crop stress resilience; M1.2 (Y2M12) Agree on commonly accepted and optimized protocols; M1.3 (Y3M3) Publication of method article on bioimaging and 3D modelling of floral meristem tissues; M1.4 (Y3M3) Online and fully operating platform for integration of –omics datasets from different crops; M1.5 (Y4M2) benchmarking and open access publication of the platform on a scientific journal
WG1 Contact: Christos Bazakos – cbazakos@elgo.gr