Microstructural and histochemical characteristics of Lycium barbarum L. fruits used in folk herbal medicine and as functional food.

Lycium barbarum L. fruits, referred to as functional food, have long been used in traditional and folk herbal medicine due to their therapeutic properties. The fruit microstructure was analysed using light, scanning and transmission electron microscopes. The distribution of bioactive compounds in drupe tissues was assessed with histochemical and fluorescence assays. The analysis of the microstructure has shown that the fruit is covered by a skin with an amorphous cuticle and a layer of amorphous epicuticular waxes on the surface. The skin is composed of a single-layered epidermis with thickened walls and one layer of hypodermis with slightly thickened periclinal walls. The pericarp cells contain different types of chromoplasts, which most often contained exhibited reticulotubules/fibrils of carotenoid pigments and phytoferritine deposits. The results of the histochemical assays demonstrated that the secondary metabolites with high phytotherapeutic importance were located in all layers of the pericarp and seeds and, specifically, in the drupe exocarp and endocarp. The phytochemicals were represented by polysaccharides (LBP), lipid compounds (carotenoids, essential oils, sesquiterpenes, steroids), polyphenols (tannins and flavonoids), and alkaloids. This study, which is the first report of the microstructure and localisation of bioactive compounds in wolfberries, is a valuable complement of phytochemical analyses and can be helpful for enhancement of the therapeutic effect of the fruit as well as preliminary assessment of the medicinal potential in the search for new pharmaceuticals. Detailed anatomical studies are crucial for exploration of determinants of fruit quality and useful for identification of diagnostic taxonomic traits.

Transfer of transformed chloroplasts from Nicotiana tabacum to the Lycium barbarum plants.

Plastid transformation is an attractive technology for obtaining crop plants with new useful characteristics and for fundamental researches of plastid functioning and nuclear-plastid interaction. The aim of our experiments was to obtain plants with Lycium barbarum nucleus and transformed Nicotiana tabacum plastids. Plastome of previously engineered transplastomic tobacco plants contains reporter uidA gene and selective aadA gene that confers resistance to antibiotics spectinomycin and streptomycin. Asymmetric somatic hybridization was performed for transferring transformed tobacco plastids from transplastomic tobacco plants into recipient L. barbarum wild type plants. Hybrid L. barbarum plants containing transformed tobacco plastome with active aadA and uidA genes were obtained as a result of the experiments. The work shows the possibility of obtaining transplastomic plants by transferring the transformed plastids to remote species by using somatic hybridization technology. The developed technique is especially effective for obtaining transplastomic plants that have low regeneration and transformation ability.

[Ultracytochemical localization of calcium and ATPase activity on the 2,4-D induced somatic embryogenesis of Lycium barbarum L].

In order to research the function mechanism of the 2,4-D during the development of plant somatic embryogenesis, we studied its function mechanism and relationship with the space-time distributing of Ca2+ content and ATPase activity on somatic embryogenesis of Lycium barbarum L. The possible effects on 2,4-dichlorophenoxyacetic acid (2,4-D) induced somatic embryogenesis and changes of Ca2+ and ATPase active at different development period of somatic embryogenesis. The result showed: The 2,4-D was a key hormone for induced embryonic state of Lycium barbarum L. The embryonic callus and non-embryonic callus was separately obtained in the medium that contains the auxin 2,4-D and lack 2,4-D. In the present study, we have observed the Ca2+ was more abundant in the further intercellular matrix and on the cell wall at the multi-cellular stage, and Ca2+ was concentrated in the plasma membrane and vacuoles membrane during embryonic cell differentiate and division, to the globular embryo, more Ca2+ was seen in the nucleus. Afterward, it was also observed to be distributed in the thicken cell wall and intercellular matrix. At the same process, the variations of ATPase activity and Ca2+ were highly similar, ATPase activity was mainly located on the plasma membrane in early embryogenic cells. With further development, it was also observed to be distributed in endoplasm, nucleus and vacuoles, with the thickening of embryogenic cell wall, ATPase activity was found in the thickened region and the intercellular space. However, the variations of ATPase activity and Ca2+ have not clearly observed variety dynamics at the nonembryogenic callus, and with further vacuolation of nonembryogenic cell, Ca2+ content and ATPase activity gradually drop. It was indicated there was a closely relationship between the dynamics of Ca2+ and ATPase activity in somatic embryogenesis by 2,4-D induced. And the space-time distribution of Ca2+ and ATPase activity play a key role on signal transmission and the regulation of relevant gene expression.