Linnaea borealis L. var. borealis-In Vitro Cultures and Phytochemical Screening as a Dual Strategy for Its Ex Situ Conservation and a Source of Bioactive Compounds of the Rare Species.

Linnaea borealis L. (Twinflower)-a dwarf shrub in the Linnaeeae tribe of Caprifoliaceae family-is distributed across the Northern Hemisphere. By means of this study, a reliable protocol for efficient micropropagation of uniform L. borealis L. var. borealis plantlets has been provided for the first time; callus culture was also established. Different initial explants, types of cultures, media systems, and plant growth regulators in Murashige and Skoog (MS) media were tested. Agitated shoot cultures in the liquid media turned out to be the best system for the production of sustainable plant biomass. After stabilization of the callus lines, the highest growth index (c.a. 526%) was gained for callus maintained on MS enriched with picloram. TLC and UHPLC-HESI-HRMS analysis confirmed the presence of phenolic acids and flavonoids, and for the first time, the presence of iridoids and triterpenoid saponins in this species. Multiplication of L. borealis shoot culture provides renewable raw material, allowing for the assessment of the phytochemical profile, and, in the future, for the quantitative analyses and the studies of the biological activity of extracts, fractions, or isolated compounds. This is the first report on in vitro cultures of traditionally used L. borealis rare taxon and its biosynthetic potential.

[Potential distribution of the traditional Tibetan herb Pterocephalus hookeri by Maxent model].

In order to study the ecology suitability of Pterocephalus hookeri, and provide a reference for GAP planting location and regional development, the Maxent model and GIS technology were used to investigate ecology suitability regions for P. hookeri based on the distribution points collected from Chinese virtual herbarium, the references and field trips. The potential distribution areas mainly concentrated in the eastern Tibet, western Sichuan, southern Qinghai, northwest Yunnan, and southern Gansu. There were 7 major environmental factors to have obvious influence on ecology suitability distributions of P. hookeri, including altitude (contribution rate of 62%), precipitation of warmest quarter (contribution rate of 14.4%), coefficient of variation of precipitation seasonality (contribution rate of 7.2%), mean temperature of driest quarter (contribution rate of 3.5%), the electrical conductivity of top and sub-soil (contribution rate of 3%), the total exchangeable bases in the top- and subsoil (contribution rate of 2.4%) and SD of temperature seasonality (contribution rate of 2.2%). The study of the ecological suitability regionalization of P. hookeri based on Maxent model can provide scientific basis for the selection of artificial planting base and GAP planting location.

Ecological risk assessment using RAPD and distribution pattern of a rare and endangered species.

Environmental and ecological risk assessment always provide useful evidence for characterisation and conservation of the rare and endangered species, e.g. seven-son flower (Heptacodium miconioides Rehd.). Seven-son flower is a deciduous arbor species, but endangered, with a restricted distribution in the subtropical forests of China. Genetic risk assessment of 56 samples of the flower from nine main populations in Zhejiang (China) was carried out by using the RAPD analysis. This was to study the ecological characteristics, spatial distribution and genetic features of the seven-son flower communities and establish a feasible conservation plan. Twenty-one primers screened from 50 yielded 119 RAPD bands with 72 polymorphic products and 60.50% of total bands. The genetic variation was found to be partitioned mainly among rather than within populations. Percentages of genetic diversity among populations were quantified by Shannon index and the Nei's gene diversity coefficient. AMOVA also demonstrated that these relict populations were highly differentiated. The high level of population variation observed is in contrast to that expected for a primarily outcrossed woody perennial plant, and suggests that there may be a degree of inbreeding. The dendrogram constructed from genetic distances through UPGMA method based on Nei's coefficients shows two groups among nine population clusters, which is further supported by a principle components analysis (PCA) of RAPD phenotypic data. The analysis showed that the biologic characteristics and habitat fragmentation were the reasons of the great genetic variation among populations. Some strategies of its genetic diversity conservation were proposed in the fragmented habitats based on its genetic structure and its biological characteristics in this study.

Use of wastewater and compost extracts as nutrient sources for growing nursery and turfgrass species.

Nutrient salts present in liquid by-products following waste treatment are lost resources if not effectively recycled, and can cause environmental problems if improperly disposed. This research compared the growth response and mineral nutrient status of two nursery and two turfgrass species, hydroponically supplied with nutritive by-product extracts derived from anaerobically digested municipal solid waste (MSW) and aerobically composted organic wastes from the mushroom and MSW industries. Forsythia (Forsythia x intermedia 'Lynwood') and weigela (Weigela florida 'Red Prince'), and creeping bentgrass (Agrostis palustris Huds.) and Kentucky bluegrass (Poa pratensis L.), were grown in nutrient solutions/extracts prepared from: (i) half-strength Hoagland's #2 solution (HH; control), (ii) Plant Products liquid fertilizer (PP; g kg(-1): 180 N; 39 P; 224 K), (iii) spent mushroom compost (SMC), (iv) MSW compost (GMC), and (v) intra-process wastewater from the anaerobic digestion of MSW (ADW). Additional nutrient solutions (SMC-A, GMC-A, and ADW-A) were prepared by amending the original solutions with N, P, and/or K to concentrations in HH (mg L(-1): 105 N; 15 P; 118 K). Plants receiving the SMC-A extract grew best or at least as well as those in HH, PP, and the amended GMC-A and ADW-A solutions. This study indicated that, with proper amendments of N, P, K and other nutrients, water-soluble constituents derived from organic waste treatment have potential for use as supplemental nutrient sources for plant production.

Lonicera Implexa leaves bearing naturally laid eggs of the specialist herbivore Euphydryas Aurinia have dramatically greater concentrations of iridoid glycosides than other leaves.

We tested in the field the hypothesis that the specialist butterfly Euphydryas aurinia (Lepidoptera: Nymphalidae, Melitaeinae) lays eggs on leaves of Lonicera implexa (Caprifoliaceae) plants with greater iridoid concentrations. We conducted our investigations in a Mediterranean site by analyzing leaves with and without naturally laid egg clusters. There were no significant differences in iridoid glycoside concentrations between leaves from plants that did not receive eggs and the unused leaves from plants receiving eggs, a fact that would seem to indicate that E. aurinia butterflies do not choose plants for oviposition by their iridoid content. However, the leaves of L. implexa that bore egg clusters had dramatically greater (over 15-fold) concentrations of iridoid glycosides than the directly opposite leaves on the same plant. These huge foliar concentrations of iridoids (15% leaf dry weight) may provide specialist herbivores with compounds that they either sequester for their own defense or use as a means of avoiding competition for food from generalist herbivores. Nevertheless, it may still be possible that these high concentrations are detrimental to the herbivore, even if the herbivore is a specialist feeder on the plant.