• Journal of Forest and Environmental Science
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• v.32 no.3
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• pp.311-322
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• 2016

Since forest trees form the basis of forest ecosystem, their prolong subsistence is crucial for various flora and fauna. The foremost challenges to sustain the forest ecosystem comprise of the declining forest tree population accompanied with structural changes due to afforestation and exploitation of forest area, environment changes, pests, pollution, and introgressive hybridization. For ex situ conservation approach, in vitro techniques encompass basic role for conserving tree genetic resources, predominantly where natural propagules like recalcitrant seed might not be appropriate for long-term conservation. The practice includes restricted growth techniques, conventional micropropagation, production and storage of synthetic seeds, and cryopreservation. Even though these practices have been applied chiefly to herbaceous species, but recently, woody species were also focused upon. Key conceptions, challenges and techniques for forest tree seed conservation are discussed briefly in this review with special emphasis on some successful cryopreservation approaches for long-term storage.

• Korean Journal of Plant Resources
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• v.24 no.4
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• pp.438-444
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• 2011

This study was conducted to investigate temperature effect on seed germination behaviors and seed viability changes by drying periods of three Ulmus species (Ulmus davidiana var. japonica (Rehder) Nakai, U. pumila L., and U. parvifolia Jacq.) distributed in Korea. Statistical analysis showed that temperature had a significant effect on mean germination time in three Ulmus species, but its effect on germination percentage was only shown in U. parvifolia (p<0.01). U. davidiana var. japonica and U. pumila germinated relatively consistent in all temperature conditions, especially the latter showed high germination behaviors in low temperatures (10-15$^{\circ}C$). U. parvifolia germinated well in high temperature (25$^{\circ}C$), while this species rarely germinated in low temperature conditions (10-20$^{\circ}C$). The results confirm that seeds of U. pumila and U. parvifolia are orthodox seeds which can be stored under 4-8% moisture condition, and U. davidiana var. japonica seed is recalcitrant seed which is difficult to store for a long time. In conclusion, these differences of the germination behaviors for three Ulmus species seem to be adaptive regimes of their natural habitats, respectively.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.47-53
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• 2008

Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

• Journal of Korean Society of Forest Science
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• v.112 no.4
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• pp.442-450
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• 2023

This study sought to determine the desiccation tolerance and storage behavior of Sarcandra glabra seeds to assist in the establishment of an ex-situ conservation strategy for the species. The basic germination characteristics of S. glabra seeds were investigated. Subsequently, the seeds were dried to various desiccation levels to allow for analysis of their germination characteristics and seed integrity. In addition, the seeds were subjected to germination tests after being stored for different durations. The initial seed moisture content following the removal of the fleshy fruit was 32.8%, and the fresh seeds maintained their seed viability even after a decrease in the seed moisture content to 4.8%, while the germination percentage was 81.0%. The germination percentage of the seeds decreased to 55.0% and 30.0% when they were desiccated to moisture contents of 2.3% and 2.2%, respectively. By contrast, when the stored seeds were desiccated, they maintained their seed viability and vigor at a moisture content of 18.6%; however, when the moisture content decreased below 5.0%, both the seed viability and vigor significantly declined and the germination percentage fell below 20.0%. Furthermore, analysis of the seed leachate revealed that the decrease in the seed viability and vigor was associated with both increased electrical conductivity and increased concentrations of inorganic compounds such as potassium and calcium. In conclusion, the findings of this study show S. glabra seeds to possess desiccation tolerance that falls somewhere between the recalcitrant and intermediate seed types, with the observed variations depending on the degree of deterioration.

• Journal of Forest and Environmental Science
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• v.34 no.5
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• pp.388-394
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• 2018

An efficient and reproducible protocol for in vitro germination of seeds of Couroupita guianensis has been developed in the present study. Couroupita guianensis is a medicinally important tree and categorized as threatened plant species by the IUCN. The seeds were surface sterilized using 0.1% $HgCl_2$ and inoculated on MS (Murashige and Skoog) and Woody Plants (Llyod and McCown) medium. The seeds with partially removed seed coat responded better (100%) on half strength MS medium as compared to the full (51%), one-fourth strength MS (79%) and the WP medium. Half strength MS medium conjunct with $2.0mg{\cdot}L^{-1}$ indole-3 butyric acid (IBA) was found best among the different types of auxins and cytokinins tested for seed germination, as all the seeds germinated on this medium combination. The seedlings were successfully hardened in the greenhouse and restored in the field with 83% survival rate. The protocol can be used as conservation measure for large scale multiplication of this threatened forest tree species.

• Korean Journal of Plant Resources
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• v.35 no.2
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• pp.183-191
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• 2022

Ginseng (Panax ginseng C.A. Meyer) is a perennial plant and propagates by seeds, and those need after-ripening for germination. To be ready for climate change and to ensure a stable seed supply, the technique for storing seeds in short-term and long-term in large quantities is required. In this study, dehisced ginseng seeds from two locations, batch #1 and batch #2, were stored at -3.5℃ with different moisture content, and after 3, 15, and 27 months of storage, the percentage of radicle emergence and shoot emergence were measured. After 3 months, radicle emergence and shoot emergence were normal only when the seed moisture content was more than 35%, and overall, germination was higher in batch #2 than in batch #1. After 15 months, the partially dehydrated seeds, with a moisture content between 45 to 54%, showed the highest germination rates, and most of the undried seeds were spoiled and failed to germinate. Seeds with moisture content lower than 25% had poor germination, too. The partially dehydrated seeds also succeeded in germination and growth in the soil after 15 months of storage, but deteriorated after one more 1 year, too. In summary, ginseng seeds look like have temperate recalcitrant seed characteristics, and partial dehydration allows extension of seed longevity.

• Journal of Korean Society of Forest Science
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• v.104 no.1
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• pp.60-66
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• 2015

This study was conducted to determine the effects of seed storage method ($-20^{\circ}C$, $2^{\circ}C$ dry, $2^{\circ}C$ wet 30 days, $2^{\circ}C$ wet 60 days, stratification and room temperature) and shading treatment(control, 50%, 80%) on seed germination, seedling growth of endangered species, Iris dichotoma and Iris setosa. As a result, seed germination rate of I. dichotoma was the highest at 75% when seed were stored at $2^{\circ}C$ wet 60 days and then sown under non-shading condition. The seed of I. dichotoma belong to intermediate seed. Seed germination rate of I. setosa was the highest at 95% when seed were stored at $2^{\circ}C$ wet 60 days and then sown under 80% shading condition. The seed of I. setosa belong to recalcitrant seed. Seedlings of I. dichotoma and I. setosa showed not only the best seedling quality but also seedling vigor index in seed stored at $2^{\circ}C$ wet 60 days under non-shading condition, with the growth characteristics of plant height (6.4, 7.2 cm), number of leaves (3, 4), leaf width (4.6, 3.2 mm), leaf length (5.7, 6.8 cm), fresh weight (aerial/root part; 144/260, 97/153 mg), dry weight (aerial/root; 31/20, 17/17 mg) and seedling vigor index and modified seedling vigor index (13,895/9,479, 13,256/8,668). In this research, I. dichotoma and I. setosa seed were stored at $2^{\circ}C$ wet 60 days, and then sown in non-shading condition, seed germination rate was more than 75%, 90%, respectively, and production of superior quality seedlings.

• Journal of Forest and Environmental Science
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• v.37 no.4
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• pp.323-330
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• 2021

Zygotic embryo culture was performed to propagate evergreen oak, Quercus myrsinifolia, which has recalcitrant seeds and is difficult to propagate by cuttings. Zygotic embryos appeared in WPM medium after 14 days, and after 56 days, they developed into complete plants with cotyledons and roots. The medium suitable for zygotic embryo culture was 1/4 WPM medium, showing a shoot growth of 2.43 cm and root growth of 8.7 cm after 8 weeks of culture. As a result of investigating the effect of GA₃ on the growth of plants germinated from zygotic embryos through GA₃ treatment, the best growth was shown in 0.5 mg/l GA₃ treatment. The in vitro rooting and growth of IBA-treated zygotic embryo-derived plants were good in the 0.5 mg/l IBA treatment and rooting and shoot growth were not observed at higher concentrations. And the callus induction rate also increased as the concentration of IBA increased. Plants grown in vitro were transferred to a plastic pot containing artificial soil and acclimatized in a greenhouse for about 4 weeks, resulting in more than 90% survival. As a result of this study, the zygotic embryo culture method was confirmed to be effective for mass propagation of Q. myrsinifolia. The results of this study are expected to contribute significantly to the mass propagation of elite Q. myrsinifolia.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.692-697
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• 2019

Agrobacterium rhizogenes-mediated transformation of sugar beet hairy roots expressing single-chain variable fragment (scFv) was exploited to evaluate the efficacy of four antibody-based constructs for interfering with the Beet necrotic yellow vein virus infection. The scFv specific to a major coat protein of virus, p21, was targeted to various cellular compartments including the cytosol (pIC and pICC constructs), apoplast (pIA), and mitochondrion (pIM). After mechanical virus inoculation, most of the hairy root clones expressing scFv in the cytosol displayed low virus titers while the majority of transgenic hairy root clones accumulated antibody in outer membrane of mitochondria or apoplast were infected. This hairy root system provided an efficient and rapid approach to initially investigating root disease resistance like rhizomania prior to transform whole recalcitrant plants such as sugar beet.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.15-15
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• 2019

The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.