• Journal of Plant Biotechnology
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• 제36권2호
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• pp.137-143
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• 2009

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

• Journal of Plant Biotechnology
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• 제5권1호
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• pp.59-61
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• 2003

Petiole explants from 20 cultivars of Catharanthus roseus were cultured on various shoot-inducing media to assess their competence for adventitious shoot formation. After eight weeks of culture on Murashige and Skoog' s medium supplemented with 4.4 $\mu\textrm{m}$6-benzyladenine and 0.5 $\mu\textrm{m}$ $\alpha$-naphthaleneacetic acid, petiole explants from 'Cooler Icy Pink' exhibited the greatest frequency of adventitious shoot formation at 40%, which was followed by 'Little Bright Eye'. By comparing with a previous study on assessment of competence for adventitious shoot formation in hypocotyl explant cultures of various cultures, it is indicated that the relative degree of their competence among cultivars varies to the organ used for the source of explant. Excised adventitious shoots were readily rooted on half-strength MS basal medium. Regenerated plantlets were successfully transplanted to potting soil and grown to maturity in a greenhouse.

• Journal of Ginseng Research
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• 제23권2호
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• pp.74-80
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• 1999

인삼 접합자배의 자엽을 식물호르몬이 전혀 첨가되지 않은 MS기본배지에 배양한 결과 자엽의 기부에서 높은 빈도의 체세포배가 유기되었다. 체세포배의 형성 빈도는 접합자배 자엽의 성숙도에 따라서 차이가 있었는데 미숙배에서 성숙배로 진행됨에 따라 감소되는 경향을 보였다. 미숙자엽의 경우에는 표피세포의 윗 아래층들이 모두 성숙자엽의 세포보다 더 적거나 더 촘촘하였으며, 많은 세포들이 체세포배의 형성에 관여하였으나 뿌리의 형성이 어려운 다배상태로 유기되었다. 그러나 발아직적의 성숙자엽은 표피세포의 윗층만이 촘촘한 세포로 이루워 졌으며 뿌리의 형성이 가능한 체세포 단일배로 유기되었다. 이런 결과는 체세포배의 기원과 발육이 배발생시 사용한 자엽의 성숙도에 따라서 배발생에 관여한 세포들이 단일 혹은 다량상태인지에 따라서 결정된다는 것을 의미한다.

• Journal of Forest and Environmental Science
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• 제25권2호
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• pp.111-118
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• 2009

Multiple shoot induction and plant regeneration using apical bud and nodal explants of 100 year old tree of Anthocephalus cadamba, an important sacred and medicinal tree in India was achieved for the first time. Aseptic explants cultured in Murashige and Skoog (MS) medium augmented with different concentrations of BAP (0.1, 0.5, 1.0, 2.5, 5.0 and 10 mg/l), when maintained for 60 days, healthy shoots were induced in presence of BAP (1 mg/l). Lower concentrations of BAP (0.1 - 0.5 mg/l) induced only one shoot per explant. Increase in number of shoots per explant was observed in presence of higher concentrations of BAP (2.5, 5.0 and 10 mg/l). However, elongation of shoots was completely inhibited. Bud break and shoot regeneration was largely associated with seasonal factors. Apical buds cultured during June to August exhibited early bud break within two weeks of initial culture. In rest of the months, bud break and shoot regeneration was very slow irrespective of the various concentrations of BAP used in the medium. Explants sourced from three different maturity levels of shoots indicated that actively growing shoots from the mother plant with 1 - 2 nodal segments was more suitable for culture initiation than the explants collected from mature shoots at dormant stage. Regenerated shoots with 2 - 3 pairs of leaves when transferred to half strength MS medium fortified with IBA (1 mg/l), 60% of the shoots induced healthy roots, indicating the possibility of large scale micropropagation.

• Plant Biotechnology Reports
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• 제3권4호
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• pp.341-345
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• 2009

This study describes culture conditions for a plant regeneration system via a combined pathway of somatic embryogenesis and organogenesis in root explant cultures of the commercial rose cultivar 'Charming'. Root explants formed white calluses at a frequency of 30% after 6 weeks of culture on Schenk and Hildebrandt (SH) medium supplemented with $11mg\;1^{-1}$ 2,4-dichlorophenoxyacetic acid. After 6 weeks of transfer to SH medium without growth regulators, initial white calluses gave rise to globular somatic embryos at a frequency of 2.8%, which were subsequently dedifferentiated to embryonic tissues. Somatic embryos or embryonic tissues initially derived from root explants did not undergo development beyond cotyledonary stage. To produce adventitious shoots, embryonic tissues were sliced and cultured on SH medium with $0.5mg\;1^{-1}$ 6-benzyladenine. After 4 weeks of culture, 28% of embryonic tissue explants formed adventitious shoots. Regenerated shoots were rooted on half strength SH medium with $0.1mg\;1^{-1}$ ${\alpha}-naphthalaneacetic$ acid and subsequently grown to maturity. Root-derived embryonic tissues were proliferated by subculture, while retaining the capacity for shoot production for a few years.

• Journal of Plant Biotechnology
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• 제5권4호
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• pp.221-224
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• 2003

Leaf explants of Ixeris sonchifolia produced adventitious shoots at a frequency of 100% when cultured on MS medium supplemented with combinations of various concentrations of 6-benzyladenine (BA) (0.44, 4.44, or 8.87 ${\mu}M$) and 0.54 ${\mu}M$ NAA, or MS medium supplemented with 22.19 ${\mu}M$ BA and 2.69 ${\mu}M\;\alpha$-naphthaleneacetic acid (NAA) after four weeks of culture. Each explants (approximately $3{\times}6mm$) produced greater than 70 shoots at a combination of 0.44 ${\mu}M$ BA and 0.54 ${\mu}M$ NAA. Leaf explants produced shoots at a frequency of greater than 80% even at as low as 0.13 ${\mu}M$ BA as the sole growth regulator. Upon transfer to one-third strength MS with 0.54 ${\mu}M$ NAA, excised adventitious shoots were rooted at a frequency of 100%. Regenerated plantlets were transplanted to potting soil and grown to maturity in a greenhouse. The competence of I. sonchifolia for plant regeneration via organogenesis appears to be greater than the competence of tobacco, currently the best model plant for organogenesis.

• Journal of Plant Biotechnology
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• 제36권2호
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• pp.193-196
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• 2009

Pinellia koreana K-H Tae & J-H Kim is a recently discovered Korea endemic medicinal plant species whose natural habitat is rapidly destroyed by industrial development. Described in this paper are culture conditions for high frequency plant regeneration via bulblet formation from leaf explant cultures of P. koreana. Leaf explants formed white nodular structures and off-white calluses at a frequency of 91.2% when cultured on MS medium supplemented with 2 mg/L BA and 0.5 mg/L NAA. However, the frequency of white nodular structures and off-white calluses formation was slightly decreased with an increasing concentration of NAA up to 4 mg/L, where the frequency reached 31.7%. Most petiole explants did not form white nodular structures and off-white calluses except the combination treatment of 2 mg/L BA and 2 mg/L NAA. Upon transfer onto MS basal medium, over 90% of nodular structures gave rise to numerous bulblets and developed into plantlets. Plantlets regenerated from bulblets were transplanted to potting soil and grown to maturity at a survival rate of over 95% in a growth chamber. Therefore, the in vitro plant regeneration system of P. koreana obtained in this study will be useful for mass propagation and long-term preservation of genetic resources of P. koreana.

• 식물조직배양학회지
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• 제21권1호
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• pp.1-6
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• 1994

멜론의 배발생 캘러스의 유도와 식물체 재분화 시스템을 확립하기 위해서 자엽과 배축의 절편을 여러 농도의 2,4-D 와 BA가 첨가된 MS 기본배지에 치상하여 관찰한 결과, 초기배양에서는 4 mg/L 2,4-D와 1 mg/L BA 그리고 2 mg/L 2,4-D+O.1 mg/L BA가 첨가된 배지에서 연한 노란색의 배발생 캘러스가 형성되었고, 위의 조건에서 형성된 캘러스로부터 배발생 캘러스는 0.5와 1 mg/L 2,4-D가 첨가된 동일배지에서 약 75%가 형성되었다. 그리고 생장조절제를 첨가하지 않은 기본배지에서 대부분의 체세포배는 식물체로 재분화되었으며, 두개의 자엽을 갖춘 정상적인 체세포배보다 비정상적인 나팔형의 자엽을 가진 것이 3배 높은 빈도로 나타났다. 이러한 체세포배들은 MS 기본배지에서 식물체로 발아되었으며 생육상에서 생장하여 견실한 열매를 맺었다.

• Journal of Plant Biotechnology
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• 제34권2호
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• pp.139-144
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• 2007

'정상' 배추의 배축절편을 선발마커로서 paromomycin 저항 성유전자를 갖고 있는 pPTN290으로 각각 형질전환된 EHA101, LBA4404, GV3101균주와 공동배양한 후 갤러스유도배지에서 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 각각 순차적으로 유도하였다. 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 가각 순차적으로 유도하였다. 형질전환캘러스 형성은 Agrobacterium균주에 따라 차이가 있었으며, 특히 EHA101균주에 공동배양된 배축절편으로부터 최대 6.1%까지 얻어졌다. 또한 각각의 형질전환캘러스 클론으로부터 형질전환 부정근과 신초 발생은 EHA101균주에서 60.7%와 38.2%, LBA4404에서 8.3%와 0%, GV3101에서 20.5%와 85.7%까지 각각 얻을 수 있었다. 형질전환식물체는 특별한 형태적 이상 없이 온실에서 정상적으로 자라 $T_{2}$종자를 얻을 수 있었다. GUS방법으로 7개의 후대 유식물체를 분석한 결과 gus유전자가 안정적으로 발현하고 있음을 확인하였고, 배추 genome에 single 또는 multiple copy로 전달되고 있음을 추측할 수 있었다.