Protein composition, chlorophyll, carotenoids, and cyanide content of cassava leaves (Manihot esculenta Crantz) as influenced by cultivar, plant age, and leaf position

The versatile use of carotenoids in feed, food, cosmetic and pharmaceutical industries has emphasized the optimization of extraction methods to obtain the highest recovery. The choice of method for carotenoid extraction from food matrices is crucial, owing to the presence of diverse carotenoids with varied levels of polarity, and the presence of various physical and chemical barriers in the food matrices. This review highlights the theoretical aspects and recent developments of various conventional and nonconventional methods used for the extraction of carotenoids, including ultrasound-assisted extraction (UAE), pressurized liquid extraction (PLE), and supercritical fluid extraction (SFE). Recent applications of non-toxic and environmentally safe solvents (green solvents) and ionic liquids (IL) for carotenoid extraction are also described. Additionally, future research challenges in the context of carotenoids extractions are also identified.

Transgenic cassava (Manihot esculenta Crantz, cv MCol22) plants with a 92% reduction in cyanogenic glucoside content in tubers and acyanogenic (<1% of wild type) leaves were obtained by RNA interference to block expression of CYP79D1 and CYP79D2, the two paralogous genes encoding the first committed enzymes in linamarin and lotaustralin synthesis. About 180 independent lines with acyanogenic (<1% of wild type) leaves were obtained. Only a few of these were depleted with respect to cyanogenic glucoside content in tubers. In agreement with this observation, girdling experiments demonstrated that cyanogenic glucosides are synthesized in the shoot apex and transported to the root, resulting in a negative concentration gradient basipetal in the plant with the concentration of cyanogenic glucosides being highest in the shoot apex and the petiole of the first unfolded leaf. Supply of nitrogen increased the cyanogenic glucoside concentration in the shoot apex. In situ polymerase chain reaction studies demonstrated that CYP79D1 and CYP79D2 were preferentially expressed in leaf mesophyll cells positioned adjacent to the epidermis. In young petioles, preferential expression was observed in the epidermis, in the two first cortex cell layers, and in the endodermis together with pericycle cells and specific parenchymatic cells around the laticifers. These data demonstrate that it is possible to drastically reduce the linamarin and lotaustralin content in cassava tubers by blockage of cyanogenic glucoside synthesis in leaves and petioles. The reduced flux to the roots of reduced nitrogen in the form of cyanogenic glucosides did not prevent tuber formation.