• Journal of Plant Biotechnology
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• v.49 no.4
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• pp.331-338
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• 2022

Sunflower (Helianthus annuus L.) protoplasts were isolated from seven-day-old etiolated hypocotyls of 10 A line and four-week-old fully expanded young leaves of PI 441983 line in vitro seedlings using an enzymatic method. Purified protoplasts were collected by filtration and floatation in sucrose solution. Semi-solid protoplast culture was performed using the L4 regeneration protocol with various culture media and plating densities to achieve the highest efficiencies for protoplast culture of hypocotyl and mesophyll protoplasts of 10 A and PI 441983 lines, respectively. The concentrations in liquid L'4M medium and different plating densities were evaluated in two types of cytokinins, the adenine-type 6-benzyladenine (BA) and the phenylurea-type thidiazuron (TDZ). The highest colony formation was achieved in both sunflower lines when 0.5 mgL^-1 BA and 0.5 mgL^-1 TDZ were applied with a high plating density (3 × 10⁵ protoplasts mL^-1). These conditions led to 38.45% and 39.40% colony formation for hypocotyl protoplasts of the 10 A line and mesophyll protoplasts of the PI 441983 line, respectively. Moreover, many hypocotyl protoplast-derived colonies developed into micro-calli. In addition, superior development of both sunflower protoplasts was observed with all plating densities when BA was used in combination with TDZ. This finding will be applicable to future sunflower hybrid production via somatic hybridization.

• Journal of Plant Biology
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• v.36 no.4
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• pp.383-389
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• 1993

The effects of light and $CO_2$ on the electrophysiological characteristics of guard cells in the intact leaf and in the detached epidermis have been investigated. Guard cells in intact leaves showed the membrane hyperpolarization in response to light. The biggest induced change of the membrane potential difference (PD) in the guard cells of the intact leaf was 13 m V by light and 42 mV by $CO_2$ in Commelina communis. Similar results were obtained with Tradescantia virginiana. However, there were no changes of membrane PD in detached epidermis. In order to determine the influence of the mesophyll on the changes of membrane PD, infiltration of the mesophyll cells with photosynthetic inhibitors was performed. In CCCP infiltrated leaf discs the guard cell membrane was depolarized slightly by red-light and hyperpolarized by $CO_2$, but in leaf discs infiltrated with DCCD and DCMU the guard cell membrane was hyperpolrized by both red-light and $CO_2$ as the control leaf discs. In azide infiltrated leaf discs the guard cell membrane showed no response to light and there was a much reduced membrane hyperpolarization by $CO_2$ compared to other responses. It was likely that azide caused leaf damage and the activity of cell metabolism was decreased greatly, resulting in small membrane PD changes by $CO_2$ and no changes by redlight. Therefore, it can be suggested that red light was sensed by the mesophyll and the light induced guard cell membrane hyperpolarization was related to energy produced by cyclic-photophosphorylation, but ${CO_2}-induced$ guard cell membrane hyperpolarization was not related to photosynthesis. Alkalisation of the vacuole was observed when the intact leaf was exposed to $CO_2$, indicating that membrane hyperpolarization was mainly the result of proton efflux.efflux.

• Korean Journal of Agricultural Science
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• v.13 no.2
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• pp.157-167
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• 1986

The study was carried out to identify several factors affecting isolation and culture of cotyledone and leaf mesophyll protoplasts of cabbage (Brassica oleraceae), petsai (B.campestris subsp. pekinensis) and rape (B.napus). High viable protoplasts could be obtained when the cotyledon and the leaf mesophyll tissue of all species were treated with enzyme solution composed of 1% macerozyme 'R-10', 1.5% Onozuka 'R-10', 10% mannitol and 50,0 mM $CaCl_22H_2O$ for 4 hours. The protoplasts which obtained from the cotyledon of all species except the cabbage and the leaf mesophyll tissue of all species were divided on NN culture medium supplemented with 9.1% mannitol, 1% glucose, 1% sucrose, $1mg/{\ell}$ 2,4-D, $0.5mg/{\ell}$ NAA and $0.5mg/{\ell}$ BA. The division of the rape leaf mesophyll protoplasts were continued and led to colony.

• The Korean Journal of Ecology
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• v.7 no.3
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• pp.99-108
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• 1983

Diurnal acid fluctuation, stomatal resistance, and solar radiation with regard to the cladode orientation were investigated in Opuntia lanceolata Haw. growing at WPeolryeong-ri, Hallim-eup, Chejudo, Korea. Diurnal changes of titratable acidity showed the typical CAM pattern in all investigated cladodes. Water tissue in the cladode had the same pattern of acid fluctuation as mesophyll tissue. Stomatal resistance was low during the night, increased rapidly to be a peak right after sunrise and decreased again thereafter. The southern side of the cladode showed higher stomatal resistance than the northern side during the day time. It suggests that the stomata of the northern side opens under diffuse radiation. The amount of solar radiation varied depending upon the cladode orientation. It is thought that C4 acids move inter and intra mesophyll tissues in the cladode through the unknown pathways. RuBP carboxyulase activity in the cladode was very high at 14:00, but was not significant at 01:00. PEP-carboxylase had high activities both at 14:00 and at 01:00. The results of this study showed the possibility that O. lanceolata Haw. had the C3, C4 and CAM photosynthetic patterns under the environmental conditions at Weolryeong-ri.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.3
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• pp.216-223
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• 2003

This study was conducted to investigate the varietal differences in leaf senescence and the relationship between leaf senescence and photosynthesis during ripening stage of rice. During grain filling period, leaf senescence was evaluated by SPAD readings (an indirect indicator of chlorophyll content) for 3 rice varieties (SNU-SG1, Hwaseongbyeo, Nampungbyeo). SPAD value of flag leaf and 2nd leaf of SNU-SG1 were much higher than the other varieties and the leaves of SNU-SG1 also showed a tendency of delayed senescence as compared to the other varieties. Photosynthesis at light saturation (Pmax) of flag, 2nd and 3rd leaf in SNU-SG1 during grain filling period were much higher than Hwaseongbyeo and Nampungbyeo. The Pmax of the flag leaf in SNU-SG1 was especially higher over 20% than the other varieties. It was due to its higher mesophyll conductance and stomatal conductance as compared to the other varieties. Pmax, stomatal conductance and mesophyll conductance had positive correlation with SPAD value and nitrogen concentration of leaves. In conclusion, the stay green characteristics of SNU-SG1 would contribute to increasing the grain yield through the improved photosynthesis during grain filling.

• Journal of Plant Biology
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• v.17 no.4
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• pp.163-170
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• 1974

Leaf tissue of Glycine max Merr. was fixed in para-formaldehyde-glutarldehyde and postfixed in osmium tetroxide or postassium permanganate for electron microscopy. The origin of cytolysome-like organelles of mesophyll cell was studied and changes of fine structure of the organelles according to treating solutions such as gibberellin (GA), kinethin (KI), 2,4-dichlorophenoxy acetic acid(2, 4-D) or 2, 4-D＋GA(1mg/l, respectively) were observed. The cylolysome-like organelles differentiate in endoplasmic reticulum and plasmalemma, and they drop into vacuoles being isolated from the formers. They seem to change into myelin-like structure and to be degenerated by autodigestion. Cytolysome-like organelles involved in cell walls and vacuoles showed activity of acid phosphatase. In the group of GA and KI treatment, cytolysome-like organelles were similar to that of the control group. But in the treatmental groups of 2,4-D and 2,4-D＋GA, myelin-like structures increased in size and autodigestion of this organelles were similar to that of the control group. But in the treatmental groups of 2,4-D and 2,4-D＋GA, myelin-like structures increased in size and autodigestion of this organelle seemed to be accelerated. In the treatmental group of 2,4-D＋GA, myelin-like structures shown high electron density were observed in cytoplasm and vacuoles together.

• Journal of the Korean Society of Tobacco Science
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• v.20 no.1
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• pp.40-49
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• 1998

Protoplasts were isolated from mesophyll of tobacco(Nicotiana glauca) transformed with kanamycin-resistant gene (NPT II gene) and potato hairy root callus containing Ri plasmid of Agrobacterium rhiEogenes, and protoplasm fusion was made between the isolated protoplasts. The transgenic tobacco leaf tissue could grow on the media containing high concentrations of kanamycin, but not on the phytohormone-free media. On the other hand, the potato hairy root calli could be cultured on the phytohormone-free media but not on media containing more than 40 ㎍/ml kanamycin. In these conditions, the viability of both protoplasts were above 90%, These selection markers were used for the selection of protoplasts fused between the two, i.e. protoplast fusion was detected using selection media containing 100㎍/ml kanamycin and with no phytohormone. The mixture of 1.0% cellulase, 0.3% macerozyme, and 0.7M mannitol was best for the maximum protoplast production for tobacco, and that of 2.0% cellulase, 2.0% macerozyme, 1.0% dricelase, and 0.5M mannitol for potato. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution on the selectable medium. Cell walls were regenerated after 5 days in this medium, and colonies were alive until 4 weeks after cultural, but died after 6 weeks.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.425-432
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• 1998

Gentian plants (Gentiana spp.) showing yellow ringspot, mosaic, necrotic fleck and malformation were collected from their growing areas in Taegu, Kyungpook province and Alpine Agricultural Experiment Station, Korea. Three viruses isolated from the naturally infected gentian were identified as broad bean wilt virus (BBWV), cucumber mosaic virus (CMV) and clover yellow vein virus identified as broad bean wilt virus (CIYVV) by their host range, immunosorbent electron microscopy (ISEM) and electron microscopy. Electron microscopic examination of negatively stained preparations showed that BBWV and CMV are spherical particles of 28 nm and 30 nm in diameter, and CIYVV is filamentous particles of ca. 780 nm in length. By ISEM, BBWV was detected mainly in gentian showing yellow ringspot and mottle, CIYVV in necrotic fleck leaf of gentian and CMV in narrow and distorted mosaic leaf of gentian. BBWV and CMV are the most prevalent in the cultivated gentian. In ultrathin sections of BBWV infected tissues, large aggregates of crystalline array of virus particles and vesicular body were found in the cytoplasm and vacuole of mesophyll cells. In case of CIYVV, pinwheel- and laminated aggregate-type inclusions as well as filamentous virus particles were observed in the cytoplasm of mesophyll cells.

• Journal of The Korean Society of Grassland and Forage Science
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• v.8 no.3
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• pp.104-109
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• 1988

A field experiment was conducted in order to investigate the seasonal changes of leaf growth and related characteristics in three cultivars of orchardgrass; Potomac, Kay and Sumas. The results were summarized as follows: 1. Leaf elongation was increased in a nearly linear phase during first and third cutting stages. It was increased slowly in early 10 days to 15 days after cutting and increased rapidly thereafter during the rest cutting stages. In cultivars, Potomac was showed hlgher leaf elongation than other cultivars during all cutting stages. There was no difference of leaf width within cutting stages, but the leaf width of fall regrowth was narrow. Sumas had relatively short and wide leaves. 2. Leaf dry weight and leaf area in first cutting stage were larger than others. Leaf area was increased rapidly from 15 days after cutting and leaf $we$ was increased rapidly from 20 days over all cutting stages. The increase in leaf area and dry weight were slow down after 30 days. 3. Number of epidermal cells was increased rapidly after cutting and the rate of increase was slow down after 30 days. In a cross section of leaf tissue, the part of mesophyll was occupied with about 60% of total area and larger area than other tissues. Leaf tissue had a large vacancy at early growth period after harvest and was filled gradually with mesophyll. This result was related to the increase of leaf dry matter.

• Journal of Plant Biology
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• v.37 no.3
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• pp.371-380
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• 1994

To determine the patterns and the levels of expression of the cauliflower mosaic virus (CaMV 35S) promoter and the rice actin 1 (Act1) promoter in rice, transgenic rice plants containing CaMV 35S-$\beta$-glucuronidase (GUS) and Act1-GUS constructs were generated and examined by fluorometric and histochemical analyses. The fluorometric analysis of stably transformed calluses showed that the activity of the rice Act1 promoter was stronger than that of the CaMV 35S promoter in rice cells. In a histochemcial study of the transgenic rices, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in parenchymal cells of vascular tissues of leaves and roots and mesophyll cells of leaves. These results are similar to those of potato, a dicot plant. In contrast, rice plant transformed with Act1-GUS fusion construct revealed strong GUS activity in parenchymal cells of vascular tissue, mesophyll cells, epidermal cells, bulliform cells, guard subsidiary cells of leaves and most cells of the root, suggesting that the rice Act1 promoter is more constitutive than the CaMV 35S promoter. It was also confirmed that in both types of transgenic rice little or no staining was localized in metaxylen tracheary elements of vascular tissue from leaves or roots. These results indicate that the rice Act1 promoter can be utilized more successfully for expression of a variety of foreign gene in rice than the CaMV 35S promoter.