Indirect organogenesis for high frequency shoot regeneration of two cultivars of Sansevieria trifasciata Prain differing in fiber production.

Sansevieria trifasciata is used as an indoor plant, in traditional medicine and as a fiber source. Here we characterized fibers of two of varieties of S. trifasciata, Lorentii and Hahnii, and report a protocol for their propagation based on indirect shoot organogenesis. Structural and ribbon fibers were scattered within leaf parenchyma when viewed with confocal laser scanning microscopy. Chemical analysis of the fibers by mass spectrometry and high-performance chromatography revealed higher contents of cellulose and xylose in Lorentii than in Hahnii and significant differences for total lignin between both. A protocol for de novo shoot production was then developed using leaf explants. Time-course histological analyses showed that the first events of transdifferentiation were triggered preferentially in cells surrounding fibers and vascular bundles. Callogenesis and shoot performances were quantified for both varieties, and 2,4-D at 2 and 3 mg·L-1 yielded the best results for primary calli induction and fresh calli mass. The length, number, and mass of shoots produced did not differ significantly between the two cultivars. The fast morphogenic response of S. trifasciata to in vitro culture may be useful for mass propagation or other biotechnological purposes such as metabolite production.

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.