IPPRAS Cryobank for the Conservation of Orthodox Seeds of Rare, Endangered, Medicinal, and Ornamental Plant Species-Current Research.

Plant cryobanks play a significant role in modern science and breeding. They contribute to the recovery of lost species, the emergence of new plant varieties, and help preserve and explore the diversity of the plant world. The IPPRAS Cryobank collection is constantly supplemented with new samples, while, at the same time, the stored samples are being monitored. In order to test seed germination, seeds of Allium and Veratrum species were thawed. Rare Allium species seeds, such as A. nutans, A. schoenoprasum, and A. victorialis were stored in liquid nitrogen for 17, 19, and 30 years, respectively. Long-term cryopreservation decreased germination rates for A. nutans from 96.55 to 50.00%, for A. schoenoprasum from 72.00 to 62.75%, and for A. victorialis from 90.00 to 83.05%. Seeds of a rare medicinal species, Veratrum lobelianum, were stored in liquid nitrogen for 18 years; the seed germination rate during this storage period has been significantly decreased from 75.00 to 14.81%. V. nigrum seeds were also collected and frozen in liquid nitrogen for 3 days. Short-term cryopreservation did not result in a statistically significant change in germination rates (from 79.71 to 82.69%). The seeds of an endangered ornamental species, Cypripedium calceolus, were collected and kept frozen for 3 days. After cryopreservation, the seeds were planted on three different media, as follows: ½ MS, MS with 10% coconut milk, and BM1. On ½ MS medium, 24.98% seeds formed protocorms, while on MS medium with 10% coconut milk, this number was 10.02%, and on BM1 medium, it was 15.02%, respectively; however, after 2.5 months, all of the protocorms died. Thus, it appears that the existing protocol for seed cryopreservation of C. calceolus needs further improvement. The size, weight, and free water content (WC) of six previously cryopreserved Stipa species and three Allium species were measured. For all the Allium and Stipa species studied, we found no correlation between seed size, WC, and cryotolerance. We also found no correlation between the life form, which reflects the water requirement of the species, and cryotolerance.

Evaluation of Seed Germination of Six Rare Stipa Species following Low Temperature Stress (Cryopreservation).

Cryopreservation is one way to preserve rare, endangered species. However, during the cryopreservation process, plant cells undergo considerable stress, which may lead to cell death. In our work, orthodox Stipa seeds of six rare species were cryopreserved: S. sareptana, S. ucrainica, S. tirsa, S. dasyphylla, S. adoxa, and S. pulcherríma. Short-term cryopreservation (14 days) stimulated germination of all Stipa species studied. Prolonged cryopreservation (70 days and more) decreased the germination of all Stipa seeds except S. sareptana. The decrease in germination progressed over time as a result of the cumulative stress of cryopreservation rather than the initial stress. To stimulate germination, seeds were stratified and treated with GA3, KNO3, NaOH, and H2O2. After four years of seed cryopreservation, it was possible to obtain seedlings of all the Stipa species studied with 30 days of stratification and 180 days of germination. After five years of cryopreservation and seed treatment with 30% NaOH for one hour, the best germination was obtained in S. adoxa and S. pulcherrima. After treatment with 5% H2O2 for 20 min, the best germination was obtained in S. sareptana, S. ucrainica, and S. dasyphylla. S. sareptana seeds germinated in all the aforementioned experiments. S. sareptana has a non-deep physiological dormancy and is the most widespread and drought-tolerant Stipa species studied. The best habitat adaptation and stress tolerance correlated with this species'cryotolerance. S. sareptana was recommended for further cryopreservation, while storage protocols for the other Stipa species studied need further improvements.

Genetic Diversity in Invasive Populations of Lupinus polyphyllus Lindl. and Heracleum sosnowskyi Manden.

In our study, two aggressive-invasive species, Lupinus polyphyllus Lindl. and Heracleum sosnowskyi Manden. from Russia and Ukraine, were investigated. The success in naturalization of both species is associated with human activities, since they have been used in agriculture and floriculture and both have qualities such as environmental tolerance, high fertility and phenotypic plasticity. The purpose of this study was to determine the level of genetic diversity of both species. For Heracleum sosnowskyi Manden., genetic diversity was compared in invasive and native populations. For Lupinus polyphyllus Lindl., the genetic diversity was compared in variety, feral and invasive populations. A genetic diversity was formulated using RAPD, ISSR and REMAP. For Heracleum sosnowskyi Manden., the average genetic diversity within the invasive population was similar (0.432), but slightly less (0.502) than within the native Caucasian population. This may suggest the successful naturalization of invaders and almost complete reconstruction of their genetic diversity. For Lupinus polyphyllus Lindl., the genetic diversity for the invasive population was the highest, with an average of 0.294, while for variety, it was the lowest, with an average of 0.194. The feral population had an intermediate place with an average of 0.248, which could suggest an increase of diversity in the process of naturalization.