Biology subjects

Sanchez Lopez, J. F.

Publications and source records attributed to Sanchez Lopez, J. F..

3 recordsLinked to original sources

bioRxiv2026

Adaptive response to long-term high temperatures during the reproductive development in Arabidopsis thaliana

Increasing global temperatures and the rising frequency of heat waves pose a significant threat to plant reproduction. The reproductive phase is particularly sensitive to heat stress, yet the underlying mechanisms regulating thermotolerance during this stage remain insufficiently understood, despite significant advcances in its understanding during vegetative growth. Heat stress responses are largely controlled by heat shock factors (HSFs) and their downstream targets, including heat shock proteins (HSPs). Among these, HSP101 is essential for acquired thermotolerance and recovery from stress, while HEAT SHOCK BINDING PROTEIN (HSBP) acts as a negative regulator of HSF activity, modulating the heat shock response. Here, we investigated the impact of elevated temperature regimes on the reproductive development of Arabidopsis thaliana, with a particular focus on pollen development and fertility. Our results show that heat stress negatively affects pollen development in a dose-dependent manner, leading to reduced reproductive success. We confirmed the critical role of HSP101 in reproductive thermotolerance using the hot1-3 mutant, deficient in HSP101. Furthermore, we provide evidence that the hot1-3 mutant is tetraploid. The origin of this event is unknown, but it is tempting to speculate that disruption of heat stress responses and interference with meiotic processes may lead to whole genome duplication. Overall, this study provides new insights into the regulation of plant reproductive development under heat stress and highlights the importance of HSP101 in maintaining fertility. These findings contribute to a better understanding of plant responses to rising temperatures and may inform strategies to enhance crop resilience under climate change. Main ConclusionFlowering Arabidopsis plants adapt to long-term high temperature by shortening the flowering period and reducing their fertility. The study also demonstrated that the commonly used hot1-3 mutant is tetraploid.

plant biology↗

bioRxiv2024

Genome editing in almond: A CRISPR-based approach through hairy root transformation

Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) technology has revolutionized genome manipulation for crop enhancement, providing a powerful toolkit. However, the tissue culture and plant regeneration steps that are critical to the CRISPR/Cas editing framework are often challenging, especially in some woody plant species that exhibit substantial resistance to these procedures. To address this, we have developed an injection-based protocol for inducing hairy roots in almond (Prunus dulcis, syn. Prunus amygdalus), a species known for its recalcitrance to conventional transformation methods. Notably, the hairy root induction method also proved effective in almond x peach hybrids. To evaluate its utility for gene functional analysis, we combined the hairy root transformation system with CRISPR/Cas9 gene editing technology, targeting two transcription factor genes (ERF74 and GAI). Our efforts resulted in transformants with target knock-out, suggesting the potential of this genetic transformation technology as a valuable tool for future routine gene function studies in almond.

plant biology↗

bioRxiv2022

Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana

Tissue clearing methods eliminate the need for sectioning, thereby helping better understand the 3D organization of tissues and organs. In the past fifteen years, clearing methods have been developed to preserve endogenous fluorescent protein tags. Some of these methods (ClearSee, TDE, PEA-Clarity, etc.) were adapted to clear various plant species, with the focus on roots, leaves, shoot apical meristems, and floral parts. However, these methods have not been used in developing seeds beyond the early globular stage. Tissue clearing is problematic in post-globular seeds due to various apoplastic barriers and secondary metabolites. In this study, we compared six methods for their efficiency in clearing Arabidopsis thaliana seeds at post-globular embryonic stages. Three methods (TDE, ClearSee, and ClearSee alpha) have been already reported in plants whereas the others (fsDISCO, FAST9, and CHAPS clear) are used in this context for the first time. These methods were assessed for seed morphological changes, clearing capacity, removal of tannins, and spectral properties. We tested each method in seeds from globular to mature stages. The pros and cons of each method are listed herein. ClearSee alpha appears to be the method of choice as it preserves seed morphology and prevents tannin oxidation. However, FAST9 with 60% iohexol as a mounting medium is faster, clears better, and appears suitable for embryonic shape imaging. Our results may guide plant researchers to choose a suitable method for imaging fluorescent protein-labeled embryos in intact Arabidopsis seeds. Key messageClearSee alpha and FAST9 were optimized for imaging Arabidopsis seeds up to the torpedo stages. The methods preserve the fluorescence of reporter proteins and seed shape, allowing phenotyping embryos in intact seeds.

plant biology↗