• Applied Microscopy
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• v.39 no.2
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• pp.73-80
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• 2009

The plastid inclusion has long been known to exist in leaves of numerous plant species, especially in those of flowering plants. Among the inclusions, crystalline bodies are the most frequently distinguished structures of the foliar plastids, however, microtubules and phytoferritins are also reported occasionally. The crystalline inclusions vary in shape, and are located either in the stroma or within intrathylakoidal spaces, whereas microtubules and phytoferritins are more uniform in shape and are formed in the stroma. In crystalline structures, the composing elements exhibit a lattice pattern and/or paralleled tubules that are either bounded by membranes or exist without membrane enclosing. Other types of inclusions have not been shown to be enclosed by any membranous structures. According to the current survey, the plastid inclusion, with the exception of phytoferritins, has been shown to exhibit a crystalline or tubular pattern, and has been reported in more than 56 species of various families. Their occurrence is not restricted to any photosynthetic pathway, but is found to be randomly distributed among C-3, C-4 and CAM species, without phylogenetic relationships. The progress in plastid inclusion research reveals more information about the function and complexity, but the need for characterizing the 3-D structure of the crystalline inclusions also has been acknowledged in previous studies. A 3-D characterization would utilize tilting and tomography of serial sections with appropriate image processing that would provide valuable information on the sub-structures of the crystalline inclusions. In fact, recent studies performed on 3-D reconstruction of the plastid inclusions revealed important information about their comprising elements. In this article, the crystals and microtubules that have been reported in various types of plastids have been reviewed, with special consideration given to their possible sub-cellular function within the plastids.

• Applied Microscopy
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• v.41 no.1
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• pp.55-60
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• 2011

Reduced plants of Spirodela polyrhiza consisting only of fronds, stalks and roots form turions during dormancy. In development, mature fronds produce offspring fronds by vegetative reproduction, and turions arise laterally from the mother frond before dormancy. The turion primordium is derived from the frond, while the frond primordium forms within the turion tissue. In the present study, cellular features, especially those of the plastids, of the above four tissue types have been examined and compared using electron microscopy. Proplastids, found to be numerous in the frond and turion primordia, differentiated into chloroplasts rapidly upon growth. The proplastids were small and the thylakoidal membrane system was rudimentary, howerver the chloroplasts exhibited variation by cell type. Chloroplasts were found within cells of the frond, stalk and root tissue. The thylakoidal membrane system, which formed grana stacks, was moderately developed within frond chloroplasts, while only a few were present in those of the stalk and root cortical cells. One to two starch grains were accumulated within frond chloroplasts, but little to none were found in stalk and root cortical chloroplasts. Contrary to other types of root chloroplasts, those found in the root cap cells developed chloroplasts similar to the frond type. Unlike proplastids of the turion primordia, numerous large amyloplasts occupied most of the turion cell volume. Moreover, the turion cell produced quite large starch grain (s) within the amyloplasts. Accumulation of the starch grains continued until they occupied the most of the stroma and in some cases, individual starch grains reached up to $9.0{\mu}m$ in length. None to little, if any, thylakoidal or internal membranous systems were seldom detected in these amyloplasts. Although the degree of cellular and tissue differentiation was rather minimal within their reduced body, the functional differentiation of Spirodela polyrhiza was very efficient, as is the case in other advanced species.

• Applied Microscopy
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• v.36 no.3
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• pp.217-226
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• 2006

Image processing by ultra high voltage electron microscopy (UHVEM) and tomography has offered major contributions to research in the field of cellular ultrastructure. Furthermore, such advancements also have enabled the improved analysis of three-dimensional cellular structures in botany. In the present study. using UHVEM and tomography, we attempted to reconstruct the three-dimensional images of plastid inclusions that probably differentiate during photosynthesis. The foliar tissues were studied Primarily with the TEM and further examined with UHVEM. The spatial relationship between tubular elements and the thylakoidal membrane and/or starch grains within plastids mainly have been investigated in CAM-performing Sedum as well as in $C_4$ Salsola species. The inclusion bodies were found to occur only in early development in the former, while they were found only in mesophyll cells in the latter. The specimens were tilted every two degrees to obtain two-dimensional images with UHVEM and subsequently comparison has been made between the two types. Digital image processing was performed on the elements of the inclusion body using tilting, tomography, and IMOD program to generate and reconstruct three-dimensional images on the cellular level. In Sedum plastids, the inclusion bodies consisted of tubular elements exhibiting about 20 nm distance between elements. However, in Salsola, plastid inclusion bodies demonstrated quite different element structure, displaying pattern, and origin relative to those of the Sedum. The inclusion bodies had an integrative relationship with the starch grains in both species.

• Applied Biological Chemistry
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• v.38 no.1
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• pp.31-36
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• 1995

To investigate the mechanism for the main chain-elongation process and the possibility of putative precursors as stater units in the biosynthesis of the Monascus pigments, feeding experiments with possible $poly-{\beta}-ketide$ intermediates were carried out. Both crotonic acid and sorbic acid, especially in low concentrations, enhanced the pigment production while not increasing the dried mycelium weight appreciably. Also, it was observed that the feeding of sorbic acid and its ethyl ester was about two folds efficient in the pigment production than the feeding of crotonic acid and its ethyl ester. In addition to these acids, cinnamic acid and vinylacrylic acid were examined for their possibility as starter units. It was observed that the color of the culture fed with cinnamic acid was dominantly dark-red, but with overall decrease in the pigment production. Whey its ethyl ester was administered to the culture, however, the pigment production increased significantly. Also noted in 2D TLC study of the pigments was the increased production of red pigment and the formation of new red pigments.

• Applied Microscopy
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• v.36 no.2
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• pp.73-82
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• 2006

Major contributions has been made in cellular ultrastructure studies with the use of high voltage electron microscopy (HVEM) and tomography. Applications of HVEM, accompanied by appropriate image processing, have provided great improvements in the analysis of three-dimensional cellular structures. In the present study, structural patterns of the crystalline bodies that are distinguished in mesophyll plastids of CAM-performing Sedum rotundifolium L., have been investigated using HVEM and tomography. Tilting, and diffraction pattern analysis were performed during the investigation. The titlting was performed at ${\pm}60^{\circ}\;with\;2^{\circ}$ increments while examining serial sections ranging from 0.125 to $1{\mu}m$ in thickness. The young plastids exhibited crystalline inclusion bodies that revealed a peculiar structural pattern. They were irregular in shape and also variable in size. Their structural attributes affected the plastid morphology. The body consisted of a large number of tubular elements, often reaching up to several thousand in number. The tubular elements typically aggregated to form a fluster The elements demonstrated either a parallel or lattice arrangement depending on the sectioning angle. The distance between the elements was approximately 20nm as demonstrated by the diffraction analysis. HVEM examination of the serial sections revealed an occasional fusion or branching of elements within the inclusion bodies. Finally, a three-dimensional reconstruction of the plastid crystalline bodies has been attempted using two different image processing methods.

• Applied Microscopy
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• v.24 no.4
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• pp.1-12
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• 1994

Various plastids in leaf and bracteal tissues of Cannabis sativa L. were examined by electron microscopy. Young chloroplasts without starch grain in mesophyll cells were ellipsoidal, and osmiophilic globules within them were common in stroma. During the plastid differentiation, the mature chloroplasts in mesophyll were changed in shape depending on the numbers and sizes of starch grain in stroma. Electron-dense granular substances were occurred along the outer membrane of chloroplasts in mesophyll. Typical plastids with reticulate body were present in the glandular trichomes. Electron-grey material appeared along the surface of a plastid. A light area in reticulate body is considered to represent junction point of thylakoids.

• Proceedings of the Korean Society of Dyers and Finishers Conference
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• 2009.11a
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• pp.21-22
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• 2009

최근, 헵타메틴 시아닌색소(heptamethinecyanine)는 그 적용 범위가 넓기 때문에 많은 연구자들의 관심을 받고 있다. 특히, photo-sensitizers, dye lasers, optical recordings와 storage media 등 다양한 분야에 적용이 가능하다. 헵타메틴 시아닌색소의 주된 특징은 polymethine 사슬에 연결된 cyclohexene 고리에 의하여 근적외선 부근에서 흡수가 이뤄진다는 것이다. 근적외선 색소의 흡수 특성을 HOMO와 LUMO 에너지 전위를 사용하여, 수치화 함으로써 분자간, 분자내 상호작용을 분석 할 수 있다. 따라서, 본 실험은 헵타메틴 시아닌 색소의 치환체에 따른 전기화학적 특성을 순환 전압-전류법(Cyclic voltammetry)과 분자 모델링을 통하여 HOMO와 LUMO의 에너지 준위를 구하고, 치환체 효과가 헵타메틴 시아닌색소에 미치는 전기화학적 특성을 UV-Vis와 계산으로부터 얻어진 에너지준위를 분석하고자 한다. 본 실험에 사용된 Uv-Vis 스펙트럼 측정은 Agilent 8453 UV-Vis spectrophotometer를 사용하였고, 전기화학적 분석 방법인 순환 전압-전류법은 Versa STAT 3 (Princeton allied research in USA)를 사용하였다. 순환전압-전류법의 측정은 Acetonitrile 용액에 $TBAPF_6$ (Tetrabutylammonium hexafluorophosphate)를 전해질로 하고, Ag/$Ag^+$을 기준전극으로 사용하여 주사 속도를 50mV/s로 하여 측정 하였다. 치환체에 의한 영향을 알아보기 위하여 분자구조 최적화 모델링을 사용하였다. 3차원 분자입체 특성 및 에너지 준위 상태는 Materials studio 4.2를 사용하여 특성을 예측 하였다. 본 연구에서는, 헵타메틴 시아닌 색소의 기본 골격에 각기 다른 치환체를 치환 시켜 치환체에 의한 영향을 전기화학적인 방법인 순환 전압-전류법(Cyclic voltammetry)와 분자 모델링 방법을 사용하여, HOMO와 LUMO에너지 준위 값을 구함으로써 치환체에 의한 영향을 알아보았다. 치환체로는 Dye 1과 Dye 2로 치환된 헵타메틴 시아닌 색소를 사용하였다. 이렇게 얻어진 HOMO/LUMO 에너지 준위 값으로부터 이온화 에너지($I_p$)와 전자 친화도($E_a$) 또한 구할 수 있는데, $I_p$와 $E_a$는 분자 오비탈과 전자전이에 관련된 값들이고, 이는 계산을 통하여 얻을 수 있다. 순환 전압-전류법의 계산 방법은 봉우리 전위(peak postential)와 (onset potential)방법이 있는데, 이 계산을 통한 전위 값들이 봉우리 전위 계산 방법이 onset potential 방법에 비하여 작은 전위 값으로 나타난다. 하지만 이 두 가지 방법 모두 현재 순환 전압-전류법을 사용하여 HOMO/LUMO 에너지 준위를 측정하는 방법에 쓰이고 있으며, 어떠한 계산 방법이 더 정확하다고는 말 할 수 없지만, 본 실험 결과를 통하여 비교 분석한 결과 onset potential 계산 방법이 봉우리 전위 계산 방법에 비하여 정확하다고 판단된다. Dye 1과 Dye 2를 순환 전압-전류법으로 측정한 결과 각기 다른 전위를 나타내고 이것을 계산을 통하여 정량화하면 Dye 2가 Dye 1에 비하여 높은 전위 값을 갖음을 알 수 있는데, 이것은 ethyl 에 비하여 surful 원자의 전자공여성이 더 크다고 할 수 있다.

• Proceedings of the Materials Research Society of Korea Conference
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• 2010.05a
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• pp.57.1-57.1
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• 2010

레이저 색소 분자간의 회합을 막음으로써 높은 레이저 효율을 갖는 고체 색소 레이저를 제조하기 위해 레이저 색소인 DCM을 유-무기 하이브리드 재료에 화학적으로 결합시켰다. 수산화기를 기능기로 갖는 DCM을 합성하여 이를 솔-젤반응의 전구체로 활용하였다. 유기실란을 DCM과 함께 솔-젤반응을 시킴으로써 DCM이 결합된 유무기 하이브리드 재료를 제조하였다. DCM이 결합된 유-무기 하이브리드 재료는 유기물과 무기물의 특성을 각각 갖고 있기 때문에 용액공정을 통해 고체 색소 레이저를 단시간에 제작할 수가 있을 뿐만 아니라 열적 및 광학적으로 우수한 특성을 보여주었다. Nd:YAG 레이저를 여기광으로 이용하여 DCM이 결합된 유-무기 하이브리드 재료의 레이저 효율을 측정하여 DCM을 유기용매에 녹인 색소 레이저의 레이저 효율과 비교하였다. DCM이 결합된 유-무기 하이브리드 재료는 DCM의 농도가 높아질 수록 유기용매를 이용한 색소 레이저에 비해서 더 높은 레이저 효율을 가지는 것을 관찰하였다. 이는 DCM이 결합된 유-무기 하이브리드 재료에 각각의 DCM 분자가 실록산 망목구조와 화학적으로 결합이 되어있기 때문에 하이브리드 재료에 안정적으로 존재할 수 있게 되고 망목구조에 둘러쌓여 분자간의 회합이 방지되는 효과를 가지게 되기 때문이다.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.183-187
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• 1998

Chlorophyll mutants were produced by treating the rhizome of Cymbidium kanran with mutagen, EMS(ethyl methan sulfonate). The germination ratio of Cymbidium kanran seeds was 5.5 times higher when the seeds were treated with ultrasonic treatment for 20 minutes than untreated control. Fifty to sixty percent of the rhizomes became dark brown when they were cultured in a liquid growth medium containing 0.2% EMS for three weeks. When the dark-brown rhizomes were cultured in a solified MS medium, new rhizomes were developed from a part of the old ones. Chlorophyll mutant rhizomes were obtained from a meristem tissue by a subculturing the cuts of these new rhizomes for a year. Of the chlorophyll mutants, a zigzag-striped type of rhizome was dominant and light-yellow and albino ones were also produced. While the zigzag-striped type rhizomes were differentiated into green and striped plant, the light yellow and the white rhizomes produced yellow-striped and albino plants repectively.These results indicate that the EMS treatment on the rhizome is an effective means to induce a chlorophyll mutant. We believe that this method may be useful to produce variegated plants chlorophyll mutants from other orchids.

• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.111-116
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• 2000

Cultural conditions for the production of water-soluble pigment from mycelial culture of Cordyceps scarabaeicola KEFC-C252 and antimutagenic activity of the pigment were investigated. To obtain the maximum productivity of the pigment from mycelial culture of C. scarabaeicola KEFC-C252, the optimized medium was made with 1.5% sucrose, 2.5% yeast extract and initial pH 5.5. C. scarabaeicola KEFC-C252 was cultivated to reach the maximum concentration of the pigment at $26^{\circ}C$ for 108 hrs. C. scarabaeicola KEFC-C252 produced about 1.2 g/liter pigment under the optimized condition. The pigment was isolated from the culture filtrate by ethylacetate extraction, acidic precipitation and crystallization. The isolated pigment was scarlet hexagonal column crystal, and the color of the pigment was changed according to pH of the solution. The pigment showed violet in the alkaline water but showed red color in the acidic water. The pigment showed inhibitory activity against mutagenic activity induced by 4-nitroquinoline N-oxide. Furthermore, the pigment showed inhibitory activity against spontaneous mutation on Salmonella typhimurium TA98 and TAlOO.