The Pennsylvania Department of Conservation and Natural Resources contacted Peter Zale after fracking destroyed a population of Spiranthes casei in the Northwestern quadrant in Pennsylvania. In November of 2015, mature seeds were harvested from two counties, air-dried for six weeks, and then subjected to three experiments. The first experiment was a simple observation of sterilization time to put seeds into tissue culture using two treatments of either 3 or 10 minutes in a 10% bleach solution. The end result was that the seeds placed in the 10 minute treatment were non-viable and completely transparent. In the second experiment seeds were plated onto three different germination media (P723, M551, and K400 available from Phytotechnology Labs). Seed germination ranged from 24 to 60% and occurred on all three media only with the 3-minute treatment. In this experiment, no significant difference was noted in relation to germination and protocorm development. The third study consisted of taking germinating seedlings and putting them into a factorial experiment using two different tissue culture media (P723 and P658) and applying three different photoperiod treatments (24-hour dark, 24-hour light, 16-hour light/8 dark) for 10 months. Seedling survival and growth occurred in all treatments but seedlings on the P723 media with the 24-hour light treatment or the 16-hour light/8 dark treatment produced plants with greater fresh weight, leaf length including more and longer roots. Conclusion of the photoperiod portion of this experiment is that light is critical for seedling development. (Taylor et al. 2018)

The Pennsylvania Department of Conservation and Natural Resources contacted Peter Zale after fracking destroyed a population of Spiranthes casei in the Northwestern quadrant in Pennsylvania. In November of 2015, mature seeds were harvested from two counties, air-dried for six weeks, and then subjected to three experiments. The first experiment was a simple observation of sterilization time to put seeds into tissue culture using two treatments of either 3 or 10 minutes in a 10% bleach solution. The end result was that the seeds placed in the 10 minute treatment were non-viable and completely transparent. In the second experiment seeds were plated onto three different germination media (P723, M551, and K400 available from Phytotechnology Labs). Seed germination ranged from 24 to 60% and occurred on all three media only with the 3-minute treatment. In this experiment, no significant difference was noted in relation to germination and protocorm development. The third study consisted of taking germinating seedlings and putting them into a factorial experiment using two different tissue culture media (P723 and P658) and applying three different photoperiod treatments (24-hour dark, 24-hour light, 16-hour light/8 dark) for 10 months. Seedling survival and growth occurred in all treatments but seedlings on the P723 media with the 24-hour light treatment or the 16-hour light/8 dark treatment produced plants with greater fresh weight, leaf length including more and longer roots. Conclusion of the photoperiod portion of this experiment is that light is critical for seedling development. (Taylor et al. 2018)

The Pennsylvania Department of Conservation and Natural Resources contacted Peter Zale after fracking destroyed a population of Spiranthes casei in the Northwestern quadrant in Pennsylvania. In November of 2015, mature seeds were harvested from two counties, air-dried for six weeks, and then subjected to three experiments. The first experiment was a simple observation of sterilization time to put seeds into tissue culture using two treatments of either 3 or 10 minutes in a 10% bleach solution. The end result was that the seeds placed in the 10 minute treatment were non-viable and completely transparent. In the second experiment seeds were plated onto three different germination media (P723, M551, and K400 available from Phytotechnology Labs). Seed germination ranged from 24 to 60% and occurred on all three media only with the 3-minute treatment. In this experiment, no significant difference was noted in relation to germination and protocorm development. The third study consisted of taking germinating seedlings and putting them into a factorial experiment using two different tissue culture media (P723 and P658) and applying three different photoperiod treatments (24-hour dark, 24-hour light, 16-hour light/8 dark) for 10 months. Seedling survival and growth occurred in all treatments but seedlings on the P723 media with the 24-hour light treatment or the 16-hour light/8 dark treatment produced plants with greater fresh weight, leaf length including more and longer roots. Conclusion of the photoperiod portion of this experiment is that light is critical for seedling development. (Taylor et al. 2018)