• Applied Microscopy
• /
• v.31 no.2
• /
• pp.207-214
• /
• 2001

Anatomy and ultrastructure of the modifeid Krana pattern have been studied in succulent Salsola collina Pall. Cylindrical leaves exhibited the Salsoloid Kranz type containing two layers of peripheral chlorenchyma that surrounded the water storage cells and vascular tissues. Small veins were also peripherally arranged, but mostly embedded in the vicinity of the inner chlorenchma without the orderly arrangement of the concentric layering of bundle sheath and mesophyll cells. The current study mainly focused on the chlorenchyma tissue abutting such minor veins. The outer columnar layer exhibited features similar to the characteristics of palisade mesophyll cells, while the inner cuboid layer to the bundle sheath cells of a typical $C_4$ Kranz pattern. Cellular components of the inner chlorenchyma were centripetal and numerous, but starch-laden chloroplasts were rudimentary in the thylakoidal system. The outer chlorenchyma demonstrated normally developed chloroplasts having well-stacked thylakoids and plastoglobuli. Branched and complicated plasmodesmata frequently occurred in thick interfaces of the two layers, implying the active movement of the photosynthates between them. The present data were mostly congruent with one of the structural features of the C4 subtypes , NADP-ME type, reported in the $C_4$ pattern. The Kranz pattern encountered in this Salsola probably has been directly related to the structural modification that occurred during a functional adaptation to the $C_4$ photosynthesis.

• Korean Journal of Agricultural Science
• /
• v.12 no.1
• /
• pp.22-30
• /
• 1985

Protoplasts of Pisum sativum L. were isolated and cultured from leaf mesophyll tissue. The successful yield of protoplast was obtained in an enzyme solution of 2% 'Onozuka R-10' and 2 % 'Macerozyme R-10' contained 6mM $CaCl_2{\cdot}2H_2O$ within 4 hours. They were divided in B5 culture medium supplemented with 2mg/l kinetin, 1mg/l 2, 4-D and 0.2% Difcobacto agar. Divisions of the protoplasts were continued and led to colony formation for 1 months. The colony from protoplasts of pea mesophyll tissue was formed to callus after subculture in a medium contained macronutrients and amino acids of BII medium and micronutrients and vitamins of B5 medium, and also supplemented with 2mg/l kinetin 2mg/l NAA.

• Korean Journal of Plant Tissue Culture
• /
• v.26 no.2
• /
• pp.127-132
• /
• 1999

Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

• Korean Journal of Weed Science
• /
• v.16 no.3
• /
• pp.237-244
• /
• 1996

Propanil [N-(3,4-dichlorophenyl) propanamide] which was applied at 4,200g ai/hapostemergence 7 days after seeding or transplanting, completely reduced the growth of shoot and root of barnyardgrass at 100% under dry condition while plant height, root length and shoot fresh weight of barnyardgrass at 63, 40 and 78%, respectively under water condition. On the other hand, the herbicide did not affect the growth of shoot and root of rice grown under water condition and transplanting condition, but reduced the plant height, root length and shoot fresh weight of broadcast rice on soil at 24, 18 and 28%, respectively, under dry condition. Microscopically, the epidermal cells of treated-barnyardgrasses under both conditions were severely constricted, chloroplasts in the cells of vascular bunble sheath were partially lacked, and mesophyll cells were often ruptured, whereas those of treated-rice were not affected. Histological observations showed that propanil reduced the thickness of leaf blade of barnyardgrass under both conditions at 36-48% due to mainly reduction and constriction of mesophyll cell, while it did not affect or even increased the thickness of leaves of rice under all conditions compared to control. These results indicate that broadcast rice on soil were more injured than drilled rice in soil under dry condition, however, in the other tested conditions ricer were not affected.

• Current Research on Agriculture and Life Sciences
• /
• v.2
• /
• pp.15-22
• /
• 1984

This study was conducted to identify the enzyme treatment time, enzyme concentration and plant growth condition for isolation of potato mesopyll, it was also performed to determine the adquate sucrose molarity on purification of protoplasts and to investigate the incubation time, PEG concentration and DMSO effect for potato-petunia protoplast fusion. The results were summarized as follows: The optimal time of incubation in enzyme solution was 3 - 4 hours and high humidity and low light intensity made plants more effective to protoplast releasing enzymes. Our experimental results showed that the pectolyase Y-23 was an ideal agent for isolation from mesophyll cultured in vitro compared with macerozyme. The enzyme solution with 0.5 % macerozyme and 2 % cellulase was very effective and the purity of healthy protoplast was better in 0.4 and 0.5 M sucrose than in others. It was revealed that the rate of potato-petunia fusion according to the incubation time with PEG was effective at 30 min incubation and percentage of protoplast aggregation was increased by high molecular weight and concentration of PEG. Percentage of potato-petunia protoplast heteroplasmic aggregation was increased by 4 to 16 % in PEG 6000 compared with PEG 4000 and PEG 1500. Addition of 5 to 10 % DMSO to the PEG solution increased to the heteroplasmic aggregation of potato-petunia from 2 to 4 %.

• KSBB Journal
• /
• v.6 no.3
• /
• pp.289-297
• /
• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.27 no.4
• /
• pp.314-339
• /
• 1982

Fluctuating climate is still most important environmental constrain, although improved modem agricultural technology has succeeded to increase crop production in the world. To stabilize the food production under fluctuating weather conditions, it is very needed to obain the quantitative information of interactions between crops and climate. The main purpose of this paper is three hold. Using the JIBP-data, the dry matter accumulation of rice crops is studied in relation to weather indexes (\SigmaTa and \SigmaSt). Temperature dependence of the yield index of rice is analyzed as to air temperature and water temperature. \SigmaT$_{10}$ -fluctuations are studied using meteorological data at various stations. The possible shift of \SigmaT$_{10}$ -isopleths due to climate fluctuation is evaluated. The second interest is in the plant climate of rice crops. Using results of canopy photosynthesis, it is pointed that the canopy structure has most important implication in plant climate. Leaf-air, stomatal, and mesophyll resistances of rice crops are described in relation to weather conditions. The change in light condition and aerodynamical property of rice crops with the growth is illustrated. The energy partition is also studied at different growing stages. Third point is to show in more detail effective countermeasures against cold irrigation water and cool summer. Heat balance of warming pond and polyethylene tube as a heat exchanger is studied to make nomo-grams for evaluating the necessary area and necessary length. Effects of windbreak net on rice crops are illustrated by using experimental and simulation results.lts.

• Plant Biotechnology Reports
• /
• v.2 no.3
• /
• pp.219-226
• /
• 2008

Transgenic plants with the herbicide-resistance gene (bar gene) were obtained via organogenesis from isolated mesophyll protoplasts of Nierembergia repens after applying electroporation. Transient ${\beta}-glucuronidase$ (GUS) activity of electroporated protoplasts assayed 2 days after applying an electric pulse showed that optimum condition (transient GUS activity 319 pmol 4 MU/mg per min and plating efficiency 2.43%) for electroporation was 0.5 kV/cm in field strength and $100{\mu}F$ in capacitance. The protoplasts electroporated with the bar gene at this condition initiated formation of microcolonies on medium after 2 weeks. After 4 weeks of culture, equal volume of fresh 1/2-strength Murashige and Skoog (MS) medium containing 0.2 mg/l bialaphos was added for selection of transformed colonies. After 6 weeks of culture, growing colonies were transferred onto regeneration medium containing 1.0 mg/l bialaphos, on which they formed adventitious shoots 1-2 months after electroporation. The adventitious shoots rooted easily after transfer onto MS medium with bialaphos lacking plant-growth regulators. Transformation of these regenerants with the bar gene was confirmed by Southern analysis. Some of the transformants showed strong resistance to the application of bialaphos solution at 10.0 mg/l.

• Applied Microscopy
• /
• v.4 no.1
• /
• pp.25-33
• /
• 1974

The origin and the function of cytolysomes were studied in the mesophyll cells and the root-tip cells of Glycine max Merr. and Zea mays L. fixed by paraformaldehyde-glutaraldehyde-$OsO_4$. The cytolysome-like structures were found of three main types of configurations: multivesicular, myelin like (multilamllar) and multitubular. More complex and mixed ones were also observed. The origin of these structures seems to be initiated by invaginations or in holdings of the plasmalemma into the cell interior, and that by aggregation and convolution of endoplasmic reticulum in the cytoplasm. Invagination of the plasmalemma were found of two main types of configurations: concentric whorls of lamellar and multivesicular. The structures were also observed within vacuoles and cytoplasm. Since the structurers are widely distributed in the cells and are greatly varied in sizes and shapes. These structures originate from the plasmalemma and the cytoplasm subsequently protrudes into the vacuole, and that seem to play an important role on the formation of the autophagic vacuoles. The possible function and fate of these structures are discussed.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
• /
• v.3 no.2
• /
• pp.121-129
• /
• 1999

Photosynthesis and transpiration rates were simultaneously measured in attached sunflower leaves(Helianthus annuusL. cv. Russian Mammoth) during exposure to $NO_2$ and $O_3$ to determine the effect of mixed gan on photosynthesis and the stomatal aperture. The application of $O_3$ alone reduced both the net photosynthetic and transpiration rates. An analysis of the $CO_2$ diffusive resistances indicated that the main cause affecting photosynthesis reduction during $O_3$ exposure was not the internal gas phase of the leaf $(rCO_2^{liq})$ but rather the liquid phase or mesophyll diffusive resistance $(rCO_2^{liq})$, suggesting that there is a very concomitant relation between photosynthetic reduction and $rCO_2^{liq}$. The application of NO2 alone caused a marked reduction of the net photosynthesis yet no significant reduction of transpiration, indicating that NO2 affects the $CO_2$ fixation processes with no inluence on the stomatal aperture. A greter reduction in the photosynthesis of sunflower plants was caused by the application of $NO_2$ alone as compared to a combination of $NO_2$ and $O_3$. $NO_2$ alone reduced the photosynthetic rate by 90%, whereas a mixture of NO2 and O3 reduced it by 50%.