Exogenous preculture with sucrose and abscisic acid improves post-cryopreservation survival of eastern bracken fern gametophytes.

Cryopreservation is an important technique used in the conservation of various plant tissues. This study proposes a cryopreservation method for the long-term conservation of eastern bracken fern gametophytes (Pteridium aquilinum var. latiusculum). Encapsulation-dehydration of the gametophytes was performed, and the exogenous sucrose and abscisic acid (ABA) preculture conditions were investigated. Gametophytes are sensitive to dehydration and drying, and the following treatment conditions were applied: encapsulation by alginate containing 0.75 M sucrose, 18-h loading treatment with 0.75 M sucrose, and 6-h drying treatment. The survival rate following cryopreservation was determined. The water content of < 27.5% in the alginate beads after dehydration and drying was found to be appropriate for ensuring survival. Additionally, performing an exogenous sucrose and ABA preculture was essential before encapsulation to achieve a survival of ≥ 90%. The high stress induced by cryopreservation and exogenous preculture regulated the expression of PaSuSy, PaLEA14, and PaABI1b and the endogenous ABA content. In eastern bracken gametophytes, ABI1 appears to be a negative regulator of ABA signaling. These results indicate that the encapsulation-dehydration method is effective for the long-term conservation of eastern bracken fern gametophytes, and exogenous preculture alleviates abiotic stress and increases the survival rate.

Exogenous gibberellin can effectively and rapidly break intermediate physiological dormancy of Amsonia elliptica seeds.

Accelerated global warming is leading to the loss of plant species diversity, and ex situ preservation of seeds is becoming an increasingly important aspect of species conservation. However, information on dormancy and germination is lacking in many endangered species. Amsonia elliptica (Apocynaceae) is the only Amsonia species native to Korea, and the South Korean Ministry of Environment has designated it Class II endangered wildlife. Nevertheless, the dormancy class and the dormancy breaking method for seeds of this species for germination are not precisely known. We identified the structure of A. elliptica seeds and the causes of dormancy, which inhibits germination. In addition, we tried to develop an effective germination promotion method by testing the wet stratified condition, which breaks dormancy, and the form of gibberellin that can replace it. Fresh seeds of A. elliptica imbibe water, but the covering layers (endosperm and seed coat) inhibit germination by mechanically restricting the embryo. Initial germination tests confirmed low embryo growth potential and physiological dormancy (PD). Restriction due to the covering layer was eliminated by seed scarification, and abnormal germination was observed. After 12 weeks of cold moist stratification at 4°C, only 12% of seeds germinated. However, 68.8% of seeds subjected to 8 weeks of warm moist stratification followed by 12 weeks of cold stratification germinated, indicating that warm stratification pretreatment before cold stratification is effective in breaking dormancy. A. elliptica seeds exhibited intermediate PD. Furthermore, 61.3% of seeds soaked in 500 mg/L GA4+7 for 14 days and incubated at 25/15°C germinated. Therefore, GA4+7 rapidly broke the dormancy of A. elliptica seeds compared with warm plus cold stratification treatment, thus providing an efficient method for seedling production.

Screening of Particulate Matter Reduction Ability of 21 Indigenous Korean Evergreen Species for Indoor Use.

The formation and pollution of particulate matter (PM), a side effect of rapid industrialization and urbanization, is considered a global issue. However, various plant species are able to effectively capture and reduce atmospheric PM concentrations. We investigated the indoor growth and morphology of 21 indigenous Korean evergreen species at low light intensities to ascertain their ability to reduce PM of aerosol particles in a closed acrylic chamber. The decrease in PM mass concentration differed significantly across species, with a significant correlation (8 h; p < 0.001). The reduction in the mass concentration of PM differed with particle size and across species. The highest reduction of PM2.5 occurred after 8 h with Dryopteris lacera (86.8%), Ilex × wandoensis (84.9%), Machilus thunbergii (84.3%), and Rhododendron brachycarpum (84.0%). Reduction of PM10 after 8 h was highest with Cephalotaxus harringtonii (98.3%), I. × wandoensis (98.5%), M. thunbergii (98.5%), and R. brachycarpum (98.3%). Plant morphological characteristics (category, plant height, leaf shape, leaf area) and relative humidity were closely related to the decrease in PM mass concentration. In conclusion, our findings can be used to identify Korean plant species that can reduce PM concentration and are suitable for indoor use.

Culture types and period impact gametophyte morphogenesis and sporophyte formation of eastern bracken.

BACKGROUND: Liquid suspension culture efficiently proliferates plant cells and can be applied to ferns because it rapidly increases the fresh weight of gametophytes. This study assessed gametophyte proliferation and sporophyte production of Pteridium aquilinum var. latiusculum using a suspension culture method. RESULTS: The growth curve linear phase of gametophyte cells was confirmed between 9 and 18 days of culture, and the subculture cycle was determined to be 2 weeks. A double-strength MS medium (fresh weight, 18.0 g) containing 2% sucrose and NH4+:NO3- (120 mM, 40:80) was found to be the optimal liquid medium. Gametophytes obtained after suspension culture for 18 days did not normally form sporophytes in an ex vitro soil environment. However, this issue was resolved after changing the culture type or extending the culture period to 6 weeks. A short suspension culture period increased the fresh weight of fragmented and homogenized gametophytes but yielded numerous relatively immature gametophytes (globular forms of branching gametophytes, BG). Furthermore, differences in gametophyte morphogenesis and development were indicated by changes in endogenous phytohormone content. BG with immature development exhibited high accumulation of zeatin, jasmonic acid, and salicylic acid, and relatively low levels of abscisic acid and indole-3-acetic acid. The immature development of gametophytes directly affected sporophyte formation. CONCLUSIONS: This study maximized the advantages of liquid suspension culture using eastern bracken gametophytes and provides data to resolve any associated issues, thus facilitating efficient bracken production.

Synthetic Seed Technology Development and Production Studies for Storage, Transport, and Industrialization of Bracken Spores.

Bracken fern (Pteridium aquilinum var. latiusculum (Desv.) Underw. ex A. Heller) has long been grown industrially in South Korea. Conventional propagation methods, including planting rhizomes and in vitro seedling culture, are labor intensive and expensive, and thus not commercially suitable. We aimed to develop a system to produce synthetic seeds using fern spores (SFS). Synthetic seeds were prepared by mixing bracken spores and alginate matrix. Spore germination and gametophyte and sporophyte growth and development from SFS proceeded normally. Spore density affected gametophyte and sporophyte numbers. SFS prepared using cold (4 °C) long-term storage spores (even 7-year-old spores) could effectively form sporophytes. The highest germination was observed at 25 °C. Soaking-treated SFS successfully formed sporophytes, even after 30 days of storage at 4 °C; indeed, sporophytes formed even after five days of storage at 25 °C during transport conditions. SFS were sown in plug trays for commercial use. Young sporophytes grown from plug seedlings were greenhouse cultivated, and transplanting within eight weeks was effective for root growth and growing-point formation. Developing synthetic seeds is a feasible solution for facilitating efficient transport and the handling of small-sized fern spores; furthermore, this SFS technology provides the basis for fern seedling culture and fern spore industrialization.

An Efficient Method for In Vitro Shoot-Tip Culture and Sporophyte Production Using Selaginella martensii Spring Sporophyte.

Selaginella martensii, an evergreen perennial fern that is native to South America and New Zealand, is named "frosty fern" because of its beautiful white-colored leaves and it is used as an ornamental plant. Efficient propagation methods for this species have not been developed. We aimed to develop an efficient propagation method for S. martensii through in vitro culture. We investigated culture conditions that are suitable for shoot-tip proliferation and growth. The optimum shoot-tip culture conditions were determined while using Murashige and Skoog (MS) medium (quarter, half, full, or double strength) and macronutrients (sucrose and two nitrogen sources) at various concentrations. In MS medium, the shoot tips formed a maximum of 6.77 nodes per explant, and each node formed two new shoot tips (i.e., 26 or 64 shoot tips). When using branching segments containing an angle meristem, the shoot-to-rhizophore formation ratio could be controlled by medium supplementation with plant-growth regulators. Sporophytes that were grown from shoot tips in vitro were acclimated in ex vitro soil conditions and successfully survived in the greenhouse. Numerous shoot tips could be obtained from in vitro-grown sporophytes and be proliferated ex vitro to produce a large number of plants. This method provides a way of shortening the time that is required for producing a large stock of S. martensii planting material.