Biology subjects

Betti, C.

Publications and source records attributed to Betti, C..

3 recordsLinked to original sources

bioRxiv2025

Developing a Molecular Toolkit to ENABLE all to apply CRISPR/Cas9-based Gene Editing in planta

Gene editing, especially the CRISPR/Cas9 (Clustered Regulatory Interspaced Short Palindromic Repeats/CRISPR associated protein 9) system, has revolutionized trait development in crops. However, large parts of the world are missing out on applying CRISPR in planta. There is an obvious lack of gene editing applications in locally relevant crops in the Global South which tend to be neglected by mainstream agricultural research and development. Access barriers to these new breeding technologies need to be removed to allow the potential impact of these technologies on food security to happen. Here, we present the ENABLE(R) Gene Editing in planta toolkit, a minimal molecular toolbox allowing users to create a CRISPR knockout vector for transient or stable plant transformation in two simple cloning steps. We validate the toolkit in rice (Oryza sativa) protoplasts and in Arabidopsis thaliana plants. The ENABLE(R) kit is designed to be utilized specifically by users in the Global South who are new to CRISPR technology by providing a simple workflow, extensive accompanying protocols as well as options for low cost methods for cloning verification and gene editing verification in planta. We hope that our toolkit helps bridge the gap between the recent biotechnological advancements in plant breeding that high income countries can access and the lack of those technologies in low and middle income countries.

plant biology↗

bioRxiv2025

The proximal proteome of FLOWERING LOCUS T LIKE 1 during rice panicle development suggests cell-to-cell mobility features

Flowering is promoted by perception of favorable environmental stimuli. In rice, exposure of leaves to short days induces the differentiation of a branched inflorescence called panicle at the shoot apical meristem (SAM). Systemic communication from the leaves to the SAM is mediated by members of the phosphatidylethanolamine-binding protein (PEBP) family, including HEADING DATE 3a (Hd3a) and RICE FLOWERING LOCUS T 1 (RFT1), commonly referred to as florigens. Florigens are [~]20kDa proteins translated in leaf companion cells and loaded into the phloematic stream, through which they reach the SAM. Once in meristematic cells, they are translocated to the nucleus, where they form higher-order protein complexes that include transcription factors and drive transcriptional reprogramming of meristematic cells. The activity of Hd3a and RFT1 at the SAM is partly mediated by FLOWERING LOCUS T LIKE 1 (FTL1) encoding a florigen-like protein required to accelerate flowering and establish the branching pattern of the panicle. The photoperiodic regulatory network is largely based on protein-protein interactions (PPIs), that take place in different tissues and in response to environmental variation. Capturing the full extent of these interactions for a specific protein and in vivo is a challenging task. Here, we developed a protocol of Proximity Labeling (PL) to study the interactome of FTL1 in developing panicles. We show that FTL1 associates with, or is proximal to, several proteins implicated in vesicular trafficking, microtubule binding and transcriptional regulation. Using a transient system, we demonstrate that FTL1 can actively move between cells, similarly to Hd3a and RFT1. This study establishes a protocol to implement PL for the study of a dynamic process during rice reproductive development, identifies interactors of FTL1, and suggests novel features possibly linked to its function.

developmental biology↗

bioRxiv2024

Impact of mutations in the HEADING DATE 1 gene on transcription and cell wall composition of rice

Plants utilize environmental information to modify their developmental trajectories for optimal survival and reproduction. Over a century ago, day length (photoperiod) was identified as a major factor influencing developmental transitions, particularly the shift from vegetative to reproductive growth. In rice, exposure to day lengths shorter than a critical threshold accelerates flowering, while longer days inhibit this process. This response is mediated by HEADING DATE 1 (Hd1), a zinc finger transcription factor that is central in the photoperiodic flowering network. Hd1 acts as a repressor of flowering under long days but functions as a promoter of flowering under short days. However, the transcriptional organization of this dual function is still not fully understood. In this study, we utilized RNA-Seq to analyze the transcriptome of hd1 mutants under both long and short day conditions. We identified genes involved in the phenylpropanoid pathway that are deregulated under long days in the mutant. Quantitative profiling of cell wall components and abiotic stress assays suggest that Hd1 is involved in processes considered unrelated to flowering control. This indicates that day length perception and responses are intertwined with physiological processes beyond flowering.

plant biology↗