• Title/Summary/Keyword: Direct shoot

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Cryopreservation of in vitro-cultured Axillary Shoot Tips of Japanese Bead Tree (Melia azedarach) using Vitrification Technique

  • Yang Byeong-Hoon;Kim Hyun-Tae;Park Ju-Yong;Park Young-Goo
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.385-391
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    • 2006
  • In vitro-grown axillary buds of Melia aredarach were successfully cryopreserved by vitrification. On the MS medium supplemented with BA 1 mg/L, multiple shoots were developed within $4{\sim}5$ weeks. Plantlets of Melia azedarach were cold-hardened at $10^{\circ}C$ for a 16-hr photo-period for 6 weeks. Excised axillary shoot-tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at $25^{\circ}C$. Axillary shoot-tip meristems wert dehydrated using a highly concentrated vitrification solution (PVS2) for 60 min at $0^{\circ}C$ prior to a direct plunge into liquid nitrogen (LN). The PVS2 vitrification solution consisted of 30% glycerol (w/v), 15% ethylene glycol (w/v), 15% DMSO (w/v) in MS medium containing 0.4M sucrose. After short-term warming in a water bath at $40^{\circ}C$, the meristems were transferred into 2 ml of MS medium containing 1.2M sucrose for 15 min and then planted on solidified MS culture medium. Successfully vitrified and warmed meristems resumed growth within 2 weeks and directly developed shoots without intermediary callus formation. The survival rate of cold-hardened plantlets for 3 and 4 weeks was 90%. We did not find any difference in PCR-band patterns between control and cryopreserved plants. This method appears to be a promising technique for cryopreserving axillary shoot-tips from in vitro-grown plantlets of Medicinal plants.

Effect of Plant Growth Regulators on in vitro Micropropagation of Colored Calla Lily(Zantedeschia spp.) (유색칼라 기내 미세번식에 미치는 식물생장조절물질의 영향)

  • Lee Young Soon;Ko Jeong Ae
    • Korean Journal of Plant Resources
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    • v.18 no.1
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    • pp.154-160
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    • 2005
  • To establish rapid micropropagation through organogenesis from apices-derived callus or direct adventitious shoot of three calla lily cultivars(Zantedeschia spp, cv. Sunlight, cv. Chiante, cv. Pink Persuation) were cultured on Murashige and Skoog medium supplemented with different plant growth regulators. The formation rate of callus, organogenesis and in viかo tuber production among the three cultivars were tested. Callus was obtained from cvs. Sunlight, Chiante and Pink Persuasion; the best cultivar was Sunlight. Sunlight induced $53.3\%$ callus and Chiante had the highest rate of $56.7\%$ direct shoot regeneration on medium with 2.0 mg/L BA. Regeneration frequencies ranged from 20 to $70\%$ on medium with 2.0-3.0 mg/L BA. The highest percentage of regeneration and the greatest number of shoots were obtained on medium containing 3.0 mg/L BA in three cultivars. Cytokinins induced multiple shoot formation; 1.0 mg/L of 2ip, 5.0 mg/L of BA, and 1.0 m/L of BA induced 16, 14 and 12 multiple shoots in cvs. Sunlight, Chiante and Pink Persuasion, respectivly. 1.0 mg/L of IAA enhanced root growth in cvs. Sunlight and Chiante while cv. Pink Persuasion exhibited enhanced root growth at 2.0 mg/L of IBA. NAA, however, induced no change in root growth. The addition of 90 g/L sucrose enhanced in vitro tuber formation and following tuber expansion in cv. Sunlight, while 70 g/L of sucrose was effective in cvs. Chiante and Pink Persuasion.

Effects of Explants and Growth Regulators on Direct Somatic Embryogenesis in Liquid Culture of Scrophularia buergeriana (현삼의 액체배양에서 체세포배 형성에 대한 치상조직과 생장조절제의 영향)

  • Song, Ji-Sook;Lim, Wan-Sang;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.294-298
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    • 1998
  • The factors affecting direct somatic embryogenesis from different parts of explant in liquid culture of Scrophularia buergeriana were investigated. Direct somatic embryogenesis was dependent on the explant tissues and stem was the most efficient explant. Rapid shoot development occurred on stem after 3-week culture but roots were not developed yet. Plantlets were not formed through somatic embryogenesis after 3-week culture of petiole. Though direct somatic embryo was not observed from leaf segment culture for 3 weeks, normal plantlets were developed after 8-week culture. BA played the main role for somatic embryogenesis in liquid culture and adding of either IAA or NAA caused rather adverse effects. Culture of stem segments in MS liquid medium with BA at 0.5 mg/ l or 0.1 mg/ l was proved to be the most efficient method for producing plantlets through direct somatic embryos.

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Micropropagation through Stem, Node-bud Shoot Tip and Bulblet Scale Culture in Fritillaria thunbergii Miq. (패모의 줄기, 마디, 정단 및 자구인편 배양에 의한 기내 증식)

  • Peak, Kee-Yoeup;Yu, Kwang-Jin;Seong, Nak-Sul;Choi, In-Sick;Cho, Jin-Tae
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.2
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    • pp.154-161
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    • 1994
  • This experiment was carried out to establish micropropagation system in Fritillaria thunbergii Miq. Through the culture of bulblet scales, stems, node-buds and shoot tips with special reference to the effect of physiological age of explant and plant growth regulators on bulblet formation. Number of formed bulblets was significantly increased in node-bud or stem tissue compared to scals segments and on the medium supplemented with kinetin than BA containing medium. Optimum levels of kinetin for bulblet formation from node-bud taken from above 3 cm shoot length and stem segments excised from below 3 cm shoot length were 5.0 mg /L and $1.0{\sim}3.0\;mg$ /L kinetin, respectively. Interesting phenomenon was observed, the direct formation of bulblets from the axilliary bud of cultured explants. Bulblet forming capacity in stem tissue was depended on stem age, young stem had high regeneration ability compared to old stem taken from above 10 cm shoot length. 1.0 mg /L kinetin was optimum concentration for the formation of bulblets from old stem segments. Stem tissue taken from underground growing plant was promoted coampare to shoot tips or bulb scale segments. Optimum concentration of sucrose was $5{\sim}7%$. Summariged above results revealed that effective explant for micropropagation was stem and /or node-bud tissue excised from less than 3 cm plant height compared to those of bulb scale segments which showed high contamination after culture. Maximum multiplication rate of young stem and /or node-bud segment was about 20 times. Kinetin requirement for stimulation of bulblet formation from cultured explant depended on source of explants but favorable levels of kinetin for organogenesis ranged from 1.0 mg /L to 5.0 mg /L.

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Influence of Nitrogen, Phosphorous, Potassium and Sucrose on the Shoot and Bulb Growth of Allium victorialis var. platyphyllum (산마늘의 신초와 구근생장에 미치는 질소, 인, 칼륨 및 당의 영향)

  • Jeong, Mi-Jin;Song, Hyun-Jin;Sim, Seon-Jeong;Doh, Eun-Soo;Chang, Jun-Pok;Yang, Jae-Kyung;Yun, Chung-Weon;Lee, Cheol-Ho;Karigar, Chandrakant. S.;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.2
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    • pp.101-107
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    • 2012
  • Effects of nitrogen (N), phosphorous (P) and potassium (K) on the shoot and bulb growth of wild garlic ($Allium$ $victorialis$ var. $platyphyllum$) were studied by adopting $in$ $vitro$ culture. These macronutrients influenced the growth of both the shoot and bulb of garlic depending upon their application doses. A minimum of 3% potassium nitrate ($KNO_3$) as a source of nitrogen was found to be critical for shoot elongation while higher concentrations were inhibitory. Garlic bulb growth was profuse on the usual $KNO_3$ strength and sucrose (7%), followed by $KNO_3$ (9.4 mM) supplement. On providing 41.22 mM ammonium nitrate ($NH_4NO_3$) as nitrogen source highest shoot growth was observed while 82.45 mM $NH_4NO_3$ as a source of nitrogen supported high bulb growth. With regard to potassium a good shoot growth was observed in medium that contained 0.31 mM $KH_2PO_4$ and 3% sucrose, while bulb growth was high on 2.5 mM $KH_2PO_4$ and 7% sucrose. These experiments may thus direct the development of excellent growth conditions for the commercial production of edible wild garlic.

Propagation of Bell Pepper (Capsicum annuum var. grossum) through tissue culture

  • Senarath, WTPSK;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.131-132
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    • 2003
  • Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

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Micropropagation and RAPD Analysis of Somaclonal Variants in Lavandula spica cv. Marino (라벤다의 기내증식과 RAPD에 의한 체세포 변이체 분석)

  • Li, Xian Ri;Seong, Eun-Soo;Kim, Il-Seop;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.2
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    • pp.94-100
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    • 1999
  • To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.

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Plant Regeneration Via Direct Adventitious Roots from Free Root Segments of Ulmus davidiana Planch (당느릅나무(Ulmus davidiana Planch) 부정근 절편으로 부터 부정아 유도를 통한 식물체 생산)

  • Kim, Ji-Ah;You, Xiang-Ling;Ahn, Chang-Ho;Lee, Jae-Seon;Choi, Yong-Eui
    • Journal of Korean Society of Forest Science
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    • v.96 no.1
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    • pp.83-88
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    • 2007
  • Micropropagation of Ulmus davidiana Planch was established via adventitious shoot formation from the segments of adventitious roots. Adventitious roots were produced directly from root segments of seedlings on a 1/2 SH medium plus various concentrations of IBA. The maximum growth of adventitious roots was observed in the presence of 2.0 mg/L IBA. After the segments of adventitious roots were cultured on various cytokinins (zeatin, 2-iP, BA, kinetin) and cytokinins plus auxin (IBA), formation of adventitious shoot was investigated. Among cytokinin treated, kinetin was the most effective on both adventitious shoot induction and number of shoots. Especially, 2.0 mg/L kinetin was the best to increase adventitious shoot induction (95.8%) and a number of shoots (8.4). Adventitious shoots were rooted on 1/2 WPM medium and the plantlets were acclimated 100% on composed soil (peatmoss : vermiculite 1 : 1).

Biomass Expansion Factors for Pinus koraiensis Forests in Korea

  • Li, Xiaodong;Yi, Myong-Jong;Jeong, Mi-Jeong;Son, Yo-Whan;Park, Pil-Sun;Lee, Kyeong-Hak;Son, Yeong-Mo;Kim, Rae-Hyun
    • Journal of Korean Society of Forest Science
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    • v.99 no.5
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    • pp.693-697
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    • 2010
  • Biomass expansion factors that convert the timber volume (or dry weight) to biomass are used to estimate the forest biomass and account for the carbon budget on a national and regional scale. This study estimated the biomass conversion and expansion factors (BCEF), root to shoot ratio (R), biomass expansion factors (BEF) and ecosystem biomass expansion factor (EBEF) of Korean pine (Pinus koraiensis) forests based on direct field surveys and publications in Korea. The mean BCEF, BEF, and R was 0.6438 Mg $m^{-3}$ (n = 7, SD = 0.1286), 1.6380 (n = 27, SD = 0.1830), and 0.2653 (n = 14, SD = 0.0698), respectively. The mean EBEF, which is a simple method for estimating the understory biomass in Korean pine forest ecosystems, was 1.0218 (n = 6, SD = 0.0090). The values of the biomass expansion factors in this study estimated the Korean pine forest biomass with more precision than the default values given by the IPCC (2003, 2006).