However, the pathogenic role of TTSuV is still inconclusive, and the target cell and tissue tropism of this virus are also ambiguous. In the present study, a multi-strained pooled probe-based in situ hybridization was established to detect the nucleic acids of TTSuV1 and TTSuV2 in the tissue. The strategy of using polymerase chain reaction-derived digoxigenin-labeled MK-1775 chemical structure multi-strained pooled probe, instead of single-strained probe or oligonucleotide, was to overcome the fact of high sequence diversity among TTSuV strains

and simultaneous infection with distinct strains of TTSuV in the same animal. The cell tropism and tissue distribution were evaluated by grading system with tissues from major organs. Lymphoid tissues, including superficial inguinal, mesenteric, and hilar lymph nodes, tonsil, intestinal lamina propria of mucosa and Peyer’s patches, and sometimes spleen, generally contained higher levels of positive signals and are considered as the learn more target sites for TTSuV. Morphologically, the distribution of TTSuV-positive signals had a strong correlation with the T lymphocyte zone. T lymphocytes are, thus, speculated as the major target cells for TTSuV. (C) 2014 Elsevier B.V. All rights reserved.”
“Premise of the study: Assessing seed quality in orchids has been hindered by stringent germination

requirements. Seed quality has traditionally been assessed in orchids using in vitro or in Selleckchem KPT-8602 situ germination protocols or viability staining. However, these methods are not always well suited for rapid assessment of viability in the context of ecological studies. Methods: The potential of an ex situ protocol for seed viability assessment of orchids in ecological studies was investigated by sowing seeds of Platanthera blephariglottis on Sphagnum moss collected in the orchid’s natural habitat. Ex situ germination

results were compared with those obtained by viability staining using triphenyltetrazolium chloride (TTC), and the effect of seed testa color on staining and germination results was investigated. Key results: The ex situ protocol yielded high germination rates, with 66% of the seeds germinating after 9 wk. Depending on the seed testa color class, ex situ germination rates were about 1.4 to 2.5 times higher than viability rates determined using TTC, indicating that the TTC technique underestimated viability compared with the method using ex situ germination. The TTC estimates of viability rates were higher for seeds with dark-colored testae than for pale ones, whereas seed testa color had no effect on germination. Conclusions: Our study showed promising results for the use of ex situ germination as an alternative to previously developed protocols for seed viability assessment of orchids in ecological studies. Staining using TTC might not be well suited for this purpose, since it introduced a bias with respect to seed testa color.