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무스카리 ‘Early Giant’ 잎 절편 유래 캘루스 배양을 통한 고빈도 체세포배 발생
이향분,전수민,한증술,김창길,임기병,정재동,정미영 한국식물생명공학회 2012 식물생명공학회지 Vol.39 No.1
Using calli of Muscari armeniacum cv. ‘Early Giant’ that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 mg・L-1 auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media,somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 mg・L-1 NAA co-supplemented with various cytokinins such as N6-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 mg・L-1 picloram supplementation were sub-cultured on medium with 0.01 mg・L-1 NAA and 0.5 mg・L-1 BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.
무스카리(Muscari armeniacum ‘Early Giant’) 엽절편 조직으로부터 신초형성과 체세포 배발생에 미치는 생장조절물질의 영향
전수민,정미영,이향분,한증술,박재석,김창길,정재동 한국화훼산업육성협회 2010 화훼연구 Vol.18 No.4
본 연구는 우리나라 주요 재배종인 Muscari armeniacum ‘Early Giant’ 품종을 사용하여 엽절편체로 부터 직접적으로 신초재생과 체세포배 발생에 미치는 생장조절제의 효과를 구명하였다. 무스카리의 엽조직으 로부터 캘러스 과정을 거치지 않은 직접 신초형성은 2,4-D 0.1 mg·L-1가 함유된 배지에서 가장 좋았다. 반면, 체세포배 발생은 생장조절제를 첨가하지 않는 대조구와 IPA 0.1~1.0 mg·L-1가 함유된 농도의 배지에서 비교적 양호하였다. 무스카리의 엽조직으로부터 재생된 자구를 기외로 이식했을 때 맹아율은 모든 처리구에서 80%이상 으로 높았으며 특히 NAA 0.1mg·L-1, IPA 1.0~3.0mg·L-1 배지에서 재생된 자구의 생장이 양호하였다. This experiment was carried out in order to determine proper plant growth regulators (PGR) and their concentrations for direct shoot induction and somatic embryogenesis from leaf tissue cultures of Muscari armeniacum ‘Early Giant’. Direct shoot formation from the leaf explant culture was effective only on a halfstrength MS medium containing 0.1 mg·L-1 2,4-D, while embryogenesis was occurred on a half-strength MS medium containing 0.1~1.0 mg·L-1 IPA or without PGR. The regenerated bulblets derived from embryos or shoots were harvested and transplanted into a greenhouse. The sprouting percentage of bulblets obtained from different culture media ranged from 80 to 100% and growth of quality bulblets was enhanced when the bulblets were harvested from the medium containing 0.1 mg·L-1 NAA and 1.0~3.0 mg·L-1 IPA.
잎 절편 배양을 이용한 배 ‘신고’ (Pyrus pyrifolia cv. Niitaka)의 신초 재분화
김세희,신일섭,조강희,김대현,김현란,김기옥,이향분,도경란,천재안,황해성 한국식물생명공학회 2013 식물생명공학회지 Vol.40 No.4
Genetic manipulation of pear (Pyrus pyrifolia Nakai) breeding is still difficult due to lack of reliable regeneration system. The aim of this research is to establish shoot regeneration system from leaf explants for pear (P. pyrifolia cv. Niitaka) using various concentrations of plant growth regulators and carbon source supplemented to medium. The highest regeneration rate of about 20% was found on a medium containing 4.4 g/L of Murashige and Skoog (MS) without vitamins, Linsmaier and Skoog (LS)vitamins were added separately. Leaf explants of pear were cultured on MS medium containing 7 g/L of Daishin agar supplemented with various concentrations of NAA (0.01,0.05, 0.1, 0.5 mg/L) in combination with BA(3, 5, 10 mg/L)for shoot regeneration. In medium with 5 mg/L of BA and 0.01 mg/L of NAA, adventitious shoot regeneration rate was higher than others treated. The optimal results were observed using MS medium supplemented with 30 g/L sorbitol as carbon source on regeneration system. Sorbitol is considered better carbon source than sucrose for shoot regeneration of pear (P. pyrifolia cv. Niitaka). In order to increase of shoot regeneration in pear (P. pyrifolia cv. Niitaka), plant agar and Daishin agar used as gelling agents, Daishin agar is more efficient in shoot regeneration.
포도 (Vitis labrusca L.)의 직접 재분화 방법을 이용한 식물체 재분화와 형질전환
김세희,신일섭,조강희,김대현,김현란,김정희,임선형,김기옥,이향분,도경란,황해성 한국식물생명공학회 2013 식물생명공학회지 Vol.40 No.4
Efficient regeneration methods and transformation system are a priority for successful application of genetic engineering to vegetative propagated plants such as grape (Vitis labrusca L.). This research is to establish shoot regeneration system from plant explants for ‘Campbell Early’, ‘Tamnara’, ‘Heukgoosul’, ‘Heukbosek’ using two types of plant growth regulators supplemented to medium. The highest adventitious shoot regeneration rate of 5% was achieved on a medium containing of Murashige and Skoog (MS) inorganic salts and Linsmaier and Skoog (LS) vitamins, 2 mg/L of TDZ and 0.1 mg/L of IBA. Leaf tissue of ‘Campbell Early’, was co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, gus gene as reporter gene and resistance to kanamycin as selective agent, the other Agrobacterium strains, GV3101 containing the vector pB7 WG2D carrying with mPAP1-D gene. mPAP1-D is a regulatory genes of the anthocyanin biosynthetic pathway. ‘Campbell Early’ harboring mPAP1-D gene was readily able to be selected by red color due to anthocyanin accumulation in the transformed shoot. These results might be helpful for further studies to enhance the transformation efficiency in grape.