DNA methylation diversity analysis of Lycium barbarum samples from different cultivation areas based on MSAP / 中国中药杂志

Obvious epigenetic differentiation occurred on Lycium barbarum in different cultivation areas in China. To investigate the difference and change rule of DNA methylation level and pattern of L. barbarum from different cultivation areas in China, the present study employed fluorescence-assisted methylation-sensitive amplified polymorphism(MSAP) to analyze the methylation level and polymorphism of 53 genomic DNA samples from Yinchuan Plain in Ningxia, Bayannur city in Inner Mongolia, Jingyuan county and Yumen city in Gansu, Delingha city in Qinghai, and Jinghe county in Xinjiang. The MSAP technical system suitable for the methylation analysis of L. barbarum genomic DNA was established and ten pairs of selective primers were selected. Among amplified 5'-CCGG-3' methylated sites, there were 35.85% full-methylated sites and 39.88% hemi-methylated sites, showing a high degree of epigenetic differentiation. Stoichiometric analysis showed that the ecological environment was the main factor affecting the epigenetic characteristics of L. barbarum, followed by cultivated varieties. Precipitation, air temperature, and soil pH were the main ecological factors affecting DNA methylation in different areas. This study provided a theoretical basis for the analysis of the epigenetic mechanism of L. barbarum to adapt to the diffe-rent ecological environments and research ideas for the introduction, cultivation, and germplasm traceability of L. barbarum.

Genetic diversity and genetic structure evaluation of cultivated Chinese wolfberry (Lycium barbarum) based on microsatellite population genetics / 中国中药杂志

A total of 178 Chinese wolfberry individuals from 17 populations were detected by 7 pairs of SSR primers to evaluate genetic diversity and structure, using software GenALEx 6.5,NTSYS,STRUCTURE, the effects of cultivation on genetic diversity and structure were clarified aiming to find the strategies for genetic management and sustainable use. The results showed that the genetic diversity of cultivated Chinese wolfberry was low. The average number of alleles N_A, expected heterozygosity H_E, observed heterozygosity H_O, and Shannon's information index H' was 3.9, 0.443 7, 0.556 6, 0.788 1, respectively. STRUCTURE, UPGMA clustering and PCA test indicated that Chinese wolfberry varieties were severely intermixed but no differentiation among varieties. Mantel test showed no significant correlation between genetic distance and geographic distance. AMOVA analysis showed that genetic variation mainly occurred among individuals within the population(84.58%, P<0.001), and there was almost no genetic differentiation between varieties(3.63%, P<0.001) and between populations(11.79%, P<0.001). The cultivation has caused a significant decline in the genetic diversity of Chinese wolfberry, which may cause inbreeding decline. New germplasm resources should be sought from the wild to improve the existing cultivars. On the other hand, there are obvious homogenization and germplasm intermixing between cultivated varieties and populations. Meanwhile, Chinese wolfberry cultivars should be purified and prevented from flowing into the wild population, in case of causing pollution of the wild germplasm.

Toxicity of the goji berry fruit associated with artificial excipients and dried without additives / Toxicidade do fruto goji berry associado a excipientes artificiais e desidratado sem aditivos

The aim of this study was to evaluate the cytotoxic and genotoxic potential of goji berry fruit-based pharmaceutical powders obtained from three pharmaceutical laboratories. The product A was tested at concentrations of 0.012; 0.025 and 0.05 g mL-1, and B and C at concentrations 0.02; 0.04 and 0.08 g mL-1. It was also evaluated the tea of the dried goji berry fruit (non-additives) in the concentrations 0.035; 0.07 and 0.14 g mL-1 for comparison to the results obtained with powdered goji berry. Tea concentrations in the two exposure times did not cause inhibition of cell division nor cellular alterations to meristem tissues. For the industrialized goji products, all concentrations analyzed caused significant antiproliferative effect to the tissues evaluated at the shortest time of analysis. There were no significant cellular changes in tissues exposed to industrialized goji. Therefore, under the conditions of analysis, goji berry powder, at the three concentrations evaluated, was cytotoxic to root meristems.

Objetivou-se na presente pesquisa avaliar, em células meristemáticas de raízes de A. cepa, nos tempos de exposição 24 e 48 horas, o potencial citotóxico e genotóxico de produtos farmacêuticos do fruto goji berry em pó, provenientes de três laboratórios farmacêuticos. O produto A foi avaliado nas concentrações 0,012; 0,025 e 0,05 g mL-1, e B e C nas concentrações 0,02; 0,04 e 0,08 g mL-1. Avaliou-se também o chá do fruto seco de goji (não aditivado), nas concentrações 0,035; 0,07 e 0,14 g mL-1, para comparação com os resultados obtidos do fruto em pó. Verificou-se que o chá nas concentrações avaliadas, nos dois tempos de exposição estabelecidos, não ocasionaram inibição da divisão celular e nem alterações celulares aos meristemas de raízes. Para os goji industrializados, todas as concentrações analisadas causaram efeito antiproliferativo significativo aos tecidos avaliados logo no menor tempo de análise considerado. Nenhum dos produtos industrializados causou número significativo de alterações aos meristemas analisados. Assim, os goji em pó foram citotóxicos ao bioensaio utilizado por terem acarretado relevante instabilidade genética aos meristemas de raízes.

Microcutting culture and morpho-physiological changes during acclimation in two Lycium chilense cytotypes

Lycium chilense, a deciduous perennial shrub, is one of the endangered native species of Patagonia due to sheep overgrazing. Chances of recolonization by seeds are scarce due to the limited density of propagules in the soil and very specific requirements for germination. The objective was to develop an in vitro propagation protocol that would help to perform reestablishment of this species in degraded areas of the Patagonian steppe. Seeds came from two provenances with different somatic number due to differences in ploidy level. Defoliated microcuttings were planted in test tubes with different growing media and taken to a growth chamber. Rooting percentage did not differ between origins, but higher values were encountered for medium without hormones. Subcultures increased significantly rooting percentage and reduced time to rooting. The leaves from micropropagated plants were thinner, did not exhibit hairs, and had poorly developed palisade parenchyma and less epicuticular waxes. In vitro leaves had lower stomatal density and their stomata were less functional when compared to acclimated leaves. A repopulation program of Lycium chilense based on microcutting culture, specialy using subcultures, is feasible.