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.

Application of SCAR (sequence characterized amplified region) analysis to authenticate Lycium barbarum (wolfberry) and its adulterants.

Fructus Lycii (Gouqizi) is well known in Chinese herbal medicine for its restorative function of benefiting the liver and kidney, replenishing vital essence and improving eyesight. However, ten species and varieties of Lycium have benn found to be substitutes or adulterants of Lycium barbarum (wolfberry) in commercial markets in the Hong Kong Special Administrative Region and in China generally. L. barbarum cv. 'Tianjinense' and Lycium chinense var. potaninii are the most common examples. It is difficult to differentiate among the Lycium species by traditional morphological and histological analyses. An easy and reliable approach based on SCAR (sequence characterized amplified region) analysis was developed in the present study to differentiate L. barbarum from other Lycium species. Two characteristic bands of approx. 700 and 650 bp were detected on the RAPD (random amplification of polymorphic DNA) profiles generated from samples of L. barbarum and L. chinense var. potaninii using the primer OPC-7. They were isolated and sequenced. Two primer sets, based on the sequences, could amplify a single specific band in samples of L. barbarum respectively, whereas no bands were detected in samples of L. chinense var. potaninii. The results confirmed that the SCAR technique can be employed for authenticating L. barbarum and its adulterants.

[Calcium distribution in the developing anther of Lycium barbarurn L].

We used potassium antimonate to precipitate "exchangeable cellular Ca2+"-calcium that is sufficiently loosely bound to combine with antimonite, to investigate the feature of calcium distribution during anther development of Lycium barbarurn L. Before the stage of microspore mother cell, few calcium-induced precipitates were found in sporogenous cells and the somatic cells of anther wall. When microspore mother cell (MMC) preparing meiosis, calcium precipitates appeared in the cytoplasm of tapetal cells and callus wall surrounding MMC. After the meiosis of MMC,abundant calcium precipitates were accumulated in the cytoplasm of early microspores,and then in pollen wall, especially in the part of germ-pores. During the late microspore stage,a big vacuole formed and the nucleus was forced to move to peripheral region. Calcium precipitates decreased sharply and might dissolve in the large vacuole. After microspore mitosis, calcium precipitates appeared in the big vacuole of 2-cellular pollen,and then the vacuole disappeared. After that, the calcium precipitates again appeared in the cytoplasm of 2-cellular pollen, and the cytoplasm became densely and storage materials like starches accumulated inside the pollen grains. When pollen maturating,many small calcium precipitates distributed in its cytoplasm,especially in nucleus. The feature of calcium distribution in the anther of Lycium barbarurn L. means that it plays some biological roles during microspore development.

[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.