Sulfatase-Activated Fluorophores for Rapid Discrimination of Mycobacterial Species and Strains.

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A cryopreservation protocol for ex situ conservation of terrestrial orchids using asymbiotic primary and secondary (adventitious) protocorms

© 2015, The Society for In Vitro Biology. In a bid to better conserve endangered terrestrial orchids, we detail cryogenic research using a widely distributed terrestrial orchid, Caladenia latifolia, as a model species for development of cryopreservation for primary (seed generated) and secondary (adventitious) protocorms. Primary protocorm cryopreservation (using droplet vitrification) involved a number of experimental lines of inquiry: investigation of a suitable plant vitrification solution (PVS) by comparing three variants of a standard PVS (2, 3 and 4), determining the most suitable primary protocorm developmental stage for successful cryopreservation, testing the effectiveness of a preculture medium treatment prior to cryopreservation, and investigating temperature preconditioning at the preculture stage as well as different components of the recovery medium. Primary protocorms were generated using asymbiotic in vitro germination media developed by the authors specifically for the test species (half-strength MS macroelements and microelements with 5% (v/v) fresh filter sterilized coconut water). Secondary protocorms were propagated using an in vitro proliferation medium (½ MS with 5 µM a-naphthaleneacetic acid + 2 µM 6-benzylaminopurine). A modified preconditioning step was developed, involving incubation on ½ MS with 0.2 M raffinose for 48 h at 15°C instead of 20°C. The standard recovery medium (½ MS 1 µM zeatin + 0.5 µM gibberellic acid) was replaced after the first week following warming from liquid nitrogen (LN), with asymbiotic germination medium (½ MS + 5% (v/v) coconut water) for the remainder of the recovery phase. This new step increased the survival of primary protocorms from 68 to 85%. The average post-cryostorage regeneration of plants from primary protocorms increased from 17 to 48% after a 6-wk incubation. A similar protocol increased the survival of secondary protocorms from 63 to 84%. Regeneration of plants from secondary cryostored protocorms increased from 11 to 26% after 14 wk. The protocols developed here provide a useful template for advancing cryoconservation of other orchid taxa, particularly rare and threatened species

Orchid re-introductions: an evaluation of success and ecological considerations using key comparative studies from Australia

With global biodiversity in decline, there is now an urgent requirement to take ameliorative action for endangered species in the form of reintroductions. For the highly diverse orchid family, many species face imminent extinction. Successful reintroductions that result in self-sustaining populations require not only an understanding of existing threats, but an in-depth understanding of species ecology. Increasingly, translocations, ranging from re-introductions to assisted colonisation, are being adopted as recovery actions. Do these translocations mitigate threatening processes and account for the two key ecological attributes for orchid survival; pollinator and mycorrhizal presence? Here, we conducted a literature review identifying the known threats to orchid survival and their necessary mitigation strategies. Next, we evaluated the success of 74 published international orchid translocations on 66 species against a consideration of orchid ecological attributes. Lastly, we empirically tested an additional 22 previously unpublished re-introductions on 12 species undertaken since 2007 against a re-introduction process that accounts for identified threats and orchid ecological attributes. We identified habitat destruction, weed invasion, herbivory, illegal collection, pollinator decline, pathogens and climate change as critical threats to orchid survival. In our global review based on published translocations, the average survival rate, 1-year post translocation was 66 % yet only 2.8 % of studies reported natural recruitment in field sites. Although survival of translocated orchids is clearly being achieved, these programmes did not relate orchid growth and development to key ecological requirements of orchid population resilience, pollinator and mycorrhizal ecology. Ensuring pollinator and mycorrhizal presence shows that these two factors alone are key factors influencing survival and persistence in an Australian review of 22 previously unpublished orchid re-introductions. In the Australian review flowering in the year following, out-planting was observed for 81 % of the re-introductions with seed set occurring in 63 % of re-introductions within the length of the study. Recruitment was observed in 18 % of the Australian re-introduced populations indicating a degree of population resilience. As orchid re-introductions will be a major strategy for wild orchid conservation in the future, we present a framework for orchid re-introductions, including criteria for success. We recommend symbiotic propagation and, for specialised pollination syndromes, the study of pollinator interactions prior to site selection and re-introduction of plants

Smoothing of initial conditions for high order approximations in option pricing

In this article the Finite Difference method is used to solve the Black Scholes equation. A second order and fourth order accurate scheme is implemented in space and evaluated. The scheme is then tried for different initial conditions. First the discontinuous pay off function of a European Call option is used. Due to the nonsmooth charac- teristics of the chosen initial conditions both schemes show an order of two. Next, the analytical solution to the Black Scholes is used when t=T/2. In this case, with a smooth initial condition, the fourth order scheme shows an order of four. Finally, the initial nonsmooth pay off function is modified by smoothing. Also in this case, the fourth order method shows an order of convergence of four.