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.

A novel genetic engineering platform for the effective management of biological contaminants for the production of microalgae.

Microalgal cultivation that takes advantage of solar energy is one of the most cost-effective systems for the biotechnological production of biofuels, and a range of high value products, including pharmaceuticals, fertilizers and feed. However, one of the main constraints for the cultivation of microalgae is the potential contamination with biological pollutants, such as bacteria, fungi, zooplankton or other undesirable microalgae. In closed bioreactors, the control of contamination requires the sterilization of the media, containers and all materials, which increases the cost of production, whereas open pond systems severely limits the number of species that can be cultivated under extreme environmental conditions to prevent contaminations. Here, we report the metabolic engineering of Chlamydomonas reinhardtii to use phosphite as its sole phosphorus source by expressing the ptxD gene from Pseudomonas stutzeri WM88, which encodes a phosphite oxidoreductase able to oxidize phosphite into phosphate using NAD as a cofactor. Engineered C. reinhardtii lines are capable of becoming the dominant species in a mixed culture when fertilized with phosphite as a sole phosphorus source. Our results represent a new platform for the production of microalgae, potentially useful for both closed photobioreactors and open pond systems without the need for using sterile conditions nor antibiotics or herbicides to prevent contamination with biological pollutants.

Phosphite cannot be used as a phosphorus source but is non-toxic for microalgae.

Phosphorous (P) plays a critical role for all living organisms as a structural component of RNA, DNA and phospholipids. Microalgae are autotrophs organisms that have been reported to only assimilate the fully oxidized phosphate (Pi) as P source. However, there are microorganisms capable of utilizing P reduced compounds (i.e. phosphite (Phi) and hypophosphite) as a sole P source, such as bacteria and cyanobacteria. In this study, we evaluated whether microalgae, such as Chlamydomonas reinhardtii, Botryococcus braunii and Ettlia oleoabundans, are capable of using Phi as a sole P source. Our studies revealed that these three microalgae are unable to use Phi as a sole P source. We also found that when Phi is present at concentrations equal or higher than that of Pi, Phi has an inhibitory effect on C. reinhardtii growth. However, since C. reinhardtii was able to survive for a long period of cultivation in the presence of high concentrations of Phi and to recover cell division capacity after transfer to media containing Pi, we noticed that Phi is not toxic for this microalga. We propose that the inhibitory effect of Phi on C. reinhardtii growth might be caused, at least in part, by a competition between the transport of Pi and Phi.