Chloroplast Isolation in Agavaceas.

Chloroplast isolation protocols have been extensively developed for various species of plants, particularly model organisms with easily manipulable physical characteristics. However, succulent plants, such as Agave angustifolia Haw., which possess adaptations for arid environments like the Crassulacean acid metabolism (CAM) and a thicker cuticle, have received less attention, resulting in a potential knowledge gap.

Protocol for Generating Arabidopsis thaliana Cell Suspension Cultures Under Different Light Conditions.

Plant cell suspension cultures (PCSCs) are in vitro-cultured cells that can divide indefinitely in a sterile growth medium. These PCSCs can be derived from various plant tissues, such as the root, stem, leaves, or seeds, and are maintained in a suitable culture medium containing nutrients, vitamins, hormones, and other essential components necessary for their growth. PCSCs have extensive applications in biotechnology, particularly in producing pharmaceutical and chemical compounds.

Transcriptome analysis reveals molecular mechanisms underlying chloroplast biogenesis in albino Agave angustifolia plantlets.

Albino plants display partial or complete loss of photosynthetic pigments and defective thylakoid membrane development, consequently impairing plastid function and development. These distinctive attributes render albino plants excellent models for investigating chloroplast biogenesis. Despite their potential, limited exploration has been conducted regarding the molecular alterations underlying these phenotypes, extending beyond photosynthetic metabolism.

Agave angustifolia albino plantlets lose stomatal physiology function by changing the development of the stomatal complex due to a molecular disruption.

Stomatal development is regulated by signaling pathways that function in multiple cellular programs, including cell fate and cell division. However, recent studies suggest that molecular signals are affected by CO concentration, light intensity, and water pressure deficit, thereby modifying distribution patterns and stomatic density and likely other foliar features as well. Here, we show that in addition to lacking chloroplasts, the albino somaclonal variants of Agave angustifolia Haw present an irregular epidermal development and morphological abnormalities of the stomatal complex, affecting the link between the stomatal conductance, transpiration and photosynthesis, as well as the development of the stoma in the upper part of the leaves.

Detection of Histone Modifications Associated with miRNAs.

The posttranslational modifications of histones and miRNAs are key epigenetic mechanisms participating in the development, growth, and reproduction of plants. Recently, coordination between these two mechanisms has been demonstrated; each mechanism can be controlled by the other for the regulation of several biological processes. For example, the acetylation of histone H3, a key modification for chromatin remodeling and gene activation, has been linked to the actions of miRNA.

Localization of miRNAs by In Situ Hybridization in Plants Using Conventional Oligonucleotide Probes.

Among the epigenetic mechanisms studied with a greater interest in the last decade are the microRNAs (miRNAs). These small noncoding RNA sequences that are approximately 17-22 nucleotides in length play an essential role in many biological processes of various organisms, including plants. The analysis of spatiotemporal expression of miRNAs provides a better understanding of the role of these small molecules in plant development, cell differentiation, and other processes; but such analysis is also an important method for the validation of biological functions.

An efficient immunodetection method for histone modifications in plants.

Background: Epigenetic mechanisms can be highly dynamic, but the cross-talk among them and with the genome is still poorly understood. Many of these mechanisms work at different places in the cell and at different times of organism development. Covalent histone modifications are one of the most complex and studied epigenetic mechanisms involved in cellular reprogramming and development in plants.