Improved method for isolation of high-quality total RNA from Agave tequilana Weber roots.

Together with their undeniable role in the ecology of arid and semiarid ecosystems, Agave species are emerging as a model to dissect the relationships between crassulacean acid metabolism and high efficiency of light and water use, and as an energy crop for bioethanol production. Transcriptome resources from economically valuable Agaves species, such as Agave tequilana and A. salmiana, as well as hybrids for fibers, are now available, and multiple gene expression landscape analyses have been reported. Key components in molecular mechanisms underlying drought tolerance could be uncovered by analyzing gene expression patterns of roots. This study describes an efficient protocol for high-quality total RNA isolation from phenolic compounds-rich Agave roots. Our methodology involves suitable root handling and collecting in the field and using saving-time commercial kits available. RNA isolated from roots free of lignified out-layers and clean cortex showed high values of quality and integrity according to electrophoresis and microfluidics-based platform. Synthesis of long full-length cDNAs and PCR amplification tested the suitability for downstream applications of extracted RNA. The protocol was applied successfully to A. tequilana roots but can be used for other Agave species that also develop lignified epidermis/exodermis in roots.

Bark water storage capacity influences epiphytic orchid preference for host trees.

PREMISE: Of all orchid species described, 70% live on phorophytes. Trees offer a vital space with characteristics that influence the successful establishment and life cycle of orchids. Field inventory and distribution analysis suggest that phorophyte selection is biased to certain tree species that would serve as better hosts. Phorophyte bark is known as an important factor that influences this preference, but the chemical and physical properties of bark that contribute to creating a favorable space for orchids are still poorly understood. In this work, the effect of bark physical characteristics on phorophyte preference of tropical orchids was studied. METHODS: Orchids and their phorophytes were counted and identified along transects inside two natural reserves in Southeast Mexico. A rhytidome classification was used to describe the bark decoration patterns of the phorophytes. To quantify bark fissuring, we developed a new protocol based on image processing of light micrographs using free-access software. Bark topology characterization was complemented with scanning electronic microscopy. Maximum and minimum water content was also determined. RESULTS: Analyses of bark decorations and bark fissuring were not enough to explain the preference found for some tropical trees. In contrast, a positive relationship was found among water-storage capacity, bark porosity, and phorophyte preference. The host trees preferred by most orchids have bark with higher pore density and higher water retention after draining. CONCLUSIONS: Unexpectedly, the phorophytes preferred by orchids are not those with more fissured bark but those with a higher ability to retain minimum water content after draining, which is a bark property positively correlated with higher pore density. Our data indicate that the bark microenvironment, determined by topology and water storage capacity, has a pivotal role in phorophyte specificity, a key factor that affects orchid diversity and distribution in the world.