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Background: Seed viability testing is essential in plant conservation and research. Seed viability testing determines the success of ex-situ conservation efforts, such as seed banking but commonly testing protocols of orchids lack consistency and accuracy, therefore, there is a need to select an appropriate and reliable viability test, especially when conducting comparative studies. Here, we evaluated the suitability of three seed viability tests, Evans blue test (EB), Fluorescein diacetate test (FDA) and Tetrazolium test (TTC), with and without sterilization, on seeds of 20 orchid species, which included five epiphytes and fifteen terrestrials, using both fresh seeds and seeds stored at − 18 ºC for 6 to 8 years. Results: We found that sterilization and lifeform of seeds affected seed viability across all tests but the storage time was not an influential factor. Sterilization negatively affected seed viability under EB and FDA test conditions but increased the detection of viable seeds in the TTC test in both epiphytic and terrestrial species. The EB test, when administered without sterilization provided the highest viability results. Being non-enzymatic unlike TTC and FDA tests, as expected, the EB test was the most reliable with similar results between sterilized and not sterilized seeds for most epiphytic and terrestrial species as well as when compared between groups. Conclusions: The lifeform of the species and seed sterilization prior to testing are important influential factors in orchid seed viability testing. Since EB test was found to be reliable we recommend the EB test for seed viability assessment in orchids rather than the less reliable but commonly used TTC test, or the FDA test, which require more expensive and sophisticated instrumentation. Since storage time was not an influential factor in orchid seed viability testing, the recommendations of this study can be used for both fresh as well as long-term stored orchid seeds. This is helpful for research and especially for conservation measures such as seed banking. However, due to the species specificity of the bio-physiology of orchids, we call for comprehensive viability test assessment in the hyper diverse orchid family to be extended to a greater number of species to facilitate efficient conservation and research. [ABSTRACT FROM AUTHOR]

Namrata Pradhan and Xuli Fan contributed equally to this work Correction to: Botanical Studies (2022) 63:3 https://doi.org/10.1186/s40529-022-00333-0 In the publication of this article (Pradhan et al. [1]), there was an error in the current (3rd) affiliation for Francesco Martini. The ORCID ids for the following authors are: Namrata Pradhan: 0000-0001-9171-8867 Xuli Fan: 0000-0002-8194-3064 Francesco Martini: 0000-0003-0806-3930 Huayang Chen: 0000-0003-4689-812X Hong Liu: 0000-0002-7814-5512 Jiangyun Gao: 0000-0003-1541-5922 The original article (Pradhan et al. [1]) has been corrected. The publication did not contain the following information about the data availability and the ORCID ids of the six authors: Availability of data and materials Data are available at Figshare Digital Repository: https://doi.org/10.6084/m9.figshare.19195628. [Extracted from the article]