Biology subjects

Segretin, M. E.

Publications and source records attributed to Segretin, M. E..

3 recordsLinked to original sources

bioRxiv2025

Phosphoinositide-specific Phospholipase C 2 (SlPLC2) Facilitates Vesicle Formation and Modulates Immune Signaling in Tomato Phytophthora infestans Interactions

Phosphoinositide-specific phospholipase C (PI-PLC) is a signaling enzyme that hydrolyzes membrane phosphoinositide to generate lipid- and lipid-derived second messengers. In plants, PI-PLCs have been implicated in various physiological processes, including immunity. Tomato SlPLC2 was previously shown to be implicated in susceptibility to the necrotrophic fungus Botrytis cinerea. However, whether this observation extends to pathogens with different lifestyles and evolutionary origins, remains unknown. Here, we investigated SlPLC2 function during infection with Phytophthora infestans, a globally devastating oomycete and causal agent of potato and tomato late blight. CRISPR-Cas9 knockout SlPLC2 plants showed significantly reduced disease symptoms, lower pathogen biomass, and decreased sporangia production, indicating impaired colonization. Upon infection, SlPLC2 knockout lines displayed attenuated expression of salicylic acid (SA)- and jasmonic acid (JA)-responsive genes suggesting disrupted hormone signaling. Consistent with PLC role during plant defense, early immune responses such as hydrogen peroxide accumulation and callose deposition were reduced in knockout plants. At the cellular level, these plants allow fewer infection vesicles formation by P. infestans, accompanied by an increase in expression of a biotrophy-associated effector gene PiAvrblb2, suggesting impaired establishment of infection. In Nicotiana benthamiana SlPLC2-GFP localizes predominantly at the plasma membrane. Upon P. infestans inoculations, SlPLC2-GFP localizes to the membrane surrounding infection vesicles, where we also detected phosphoinositides PI4P and PI(4,5)P2. Overexpression of SlPLC2 in Nicotiana benthamiana enhanced susceptibility to P. infestans, reinforcing its positive role in colonization. Together, these findings identify SlPLC2 as a susceptibility factor associated with enhanced P. infestans colonization. StatementThis study identifies SlPLC2 as a key susceptibility factor that supports Phytophthora infestans infection by coordinating vesicle formation and early immune responses in tomato.

plant biology↗

bioRxiv2024

Production of biologically active human basic Fibroblast Growth Factor (hFGFb) using Nicotiana tabacum transplastomic plants

The use of plants as biofactories presents as an attractive technology with the potential to efficiently produce high-value human recombinant proteins in a cost-effective manner. Plastid genome transformation stands out for its possibility to accumulate recombinant proteins at elevated levels. Of particular interest are recombinant growth factors, given their applications in animal cell culture and regenerative medicine. In this study we produced recombinant human Fibroblast Growth Factor (rhFGFb), a crucial protein required for animal cell culture, in tobacco chloroplasts. We successfully generated two independent transplastomic lines that are homoplasmic and accumulate rhFGFb in their leaves. Furthermore, the produced rhFGFb demonstrated its biological activity by inducing proliferation in HEK293T cell lines. These results collectively underscore plastid genome transformation as a promising plant-based bioreactor for rhFGFb production. Main conclusionWe generated transplastomic tobacco lines that stably express a human Basic Fibroblast Growth Factor (hFGFb) in their chloroplasts stroma and purified a biologically active recombinant hFGFb.

bioengineering↗

bioRxiv2019

Chloroplasts navigate towards the pathogen interface to counteract infection by the Irish potato famine pathogen

Upon immune activation, chloroplasts switch off photosynthesis, produce anti-microbial compounds, and associate with the nucleus through tubular extensions called stromules. Although it is well-established that chloroplasts alter their position in response to light, little is known about the dynamics of chloroplasts movement in response to pathogen attack. Here, we report that chloroplasts accumulate at the pathogen interface during infection by the Irish potato famine pathogen Phytophthora infestans, associating with the specialized membrane that engulfs the pathogen haustorium. Chemical inhibition of actin polymerization reduces the accumulation of chloroplasts at the pathogen haustoria, suggesting this process is partially dependent on the actin cytoskeleton. However, chloroplast accumulation at haustoria does not necessarily rely on movement of the nucleus to this interface and is not affected by light conditions. Stromules are typically induced during infection, embracing haustoria and interconnecting chloroplasts, to form dynamic organelle clusters. We found that infection-triggered stromule formation relies on BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED KINASE 1 (BAK1) mediated surface immune signaling, whereas chloroplast repositioning towards haustoria does not. Consistent with the defense-related induction of stromules, effector mediated suppression of BAK1 mediated immune signaling reduced stromule formation during infection. On the other hand, immune recognition of the same effector stimulated stromules, presumably via a different pathway. These findings implicate chloroplasts in a polarized response upon pathogen attack and point to more complex functions of these organelles in plant-pathogen interactions.

plant biology↗