Effect of temperature and storage duration on seed quality of three Hypericum (Hypericaceae) species

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Abstract

The effect of storage temperatures and duration on long-term storage of the seeds of three Hypericum species (Hypericaceae): H. perforatum L., H. maculatum Crantz., H. hirsutum L. was studied. The seed quality was assessed by laboratory germination and germination dynamics tests before storage, after one-month cryopreservation in liquid nitrogen, and then every 3 years over 18 years of storage at 5, —20 and –196 °C (in liquid nitrogen). The development of seedlings from the seeds that have been cryopreserved or cold-stored for 9 years and longer was evaluated. In all studied species, seed quality was not affected by the short-term cryopreservation. When stored at 5 °C, the seeds of H. perforatum retained their quality for 6 years, and of H. maculatum — for 15 years. The seeds of H. perforatum and H. maculatum stored at –20 °C retained their quality up to 6 years, and at –196 °C — along the entire 18 years of monitoring. The quality of H. hirsutum seeds that were placed for long-term storage after 18 months of room storage, deteriorated significantly after 6 years of storage at 5 and –20 °C. When additional lot of H. hirsutum fresh seeds (after 5 months of room storage) were placed for long-term storage, they retained quality at 5 °C for 9 years, and at –20 °C — for 12 years (the entire period of monitoring). In liquid nitrogen, the seeds of both H. hirsutum sample lots retained their quality for a long time. After 18 years of storage, in all three species, the best sowing qualities were observed in seeds stored at –196 °C, and the lowest — at –20 °C. In seeds stored under all above-mentioned temperatures, the germination ability changed insignificantly, and the germination rate decreased under storage at –20 and 5 °C. Under long-term storage at –20 °C, the seeds retained the ability for root emergence, but the size of the seedlings decreased; in H. hirsutum the number of normally developing seedlings also decreased, which means the faster ageing of epicotyl meristem than the radicle meristem. An increase in the duration of the preliminary room storage led to an accelerated ageing of seeds stored both at 5 and at –20 °C. Storage at –196 °C significantly decreased seed ageing, regardless of the duration of the preliminary room storage. Seeds stored in liquid nitrogen are more sensitive to the swelling and germination conditions than those stored at 5 and –20 °C. The desiccation and ageing resistance of H. hirsutum seeds is lower than that of H. perforatum and H. maculatum. It is suggested to use seed recovery after short-term cryopreservation under suboptimal conditions as a test for seed resistance to desiccation and suitability for long-term storage. Recommendations for the studied species’ seed quality monitoring during their long-term storage are given.

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About the authors

G. E. Levitskaya

Institute of Cell Biophysics of the Russian Academy of Sciences

Author for correspondence.
Email: levitskaya_g@mail.ru
Russian Federation, Pushchino

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2. Fig. 1. Seed laboratory germination (%)* of three Hypericum species depending on the storage temperature and time. a — H. perforatum, b — H. maculatum, c — H. hirsutum. X-axis — storage time, years; y-axis — laboratory germination (%). Storage temperature: ▄ initial value ▄ –196 °C (liquid nitrogen) ▄ –20 °C ▄ 5 °C *– arithmetic mean for 4×50 seeds ± standard error of mean.

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3. Fig. 2. Seed germination dynamics of three Hypericum species after 18 years of storage at different temperatures. X-axis — germination time, days; y-axis — germination, %. Storage temperature: 1 — -196 °C (in liquid nitrogen), 2 — -20 °C, 3—5 °C.

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4. Fig. 3. Seedlings development of three Hypericum species after 18 years of seed storage at different temperatures. a — H. perforatum, b — H. maculatum, c — H. hirsutum. Storage temperature: 1 — -196 °C (in liquid nitrogen), 2 — -20 °C, 3—5 °C.

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5. Fig. 4. Seedlings development of Hypericum hirsutum (sample 1) after 9 years of seed storage at different temperatures followed by different rehydration methods. Storage temperature: 1 — -196 °C (in liquid nitrogen), 2 — -20 °C, 3 — 5 °C. Post-storage rehydration methods: a — humidification with water at 24 °C one hour after removal from storage; b — in air at 24 °C and relative humidity of 65% for 3 days.

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6. Fig. 5. Laboratory germination (%)* of Hypericum hirsutum seeds (sample 2) depending on the storage temperature and time. X-axis — storage time, years; y-axis — laboratory germination, %. Storage temperature: initial value, −196 °C (liquid nitrogen), −20 °C, 5 °C. * — arithmetic mean for 4×50 seeds ± standard error of mean.

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