• The Plant Pathology Journal
• /
• v.21 no.4
• /
• pp.395-401
• /
• 2005

The dlm (disease lesion mimic) mutant of soybean (Glycine max L. Merr) shows the similar lesion of a soybean disease caused by a fungus, Corynespora cassilcola. The lesion was examined at cellular and molecular level. Trypan blue staining result indicated that cell death was detectable in the entire region of leaves excluding veins when the lesions had already been developed. We found that the mesophyll cells of palisade layer in the dim mutant appeared to be wider apart from each other. The chloroplasts of the dim mutant cells contained bigger starch granules than those in normal plants. We also found that the lesion development of dlm plant was light-dependent and the starch degradation during the dark period of diurnal cycle was impaired in the mutant. Three soybean pathogenesis-related genes, PR-1a, PR-4, and PR-10, were examined for their expression patterns during the development of disease lesion mimic. The expression of all three genes was up-regulated to some extent upon the appearance of the disease lesion mimic. Although the exact function of DLM protein remains elusive, our data would provide some insight into mechanism underling the cell death associated with the dim mutation.

• Molecules and Cells
• /
• v.27 no.1
• /
• pp.67-74
• /
• 2009

Autophagy degrades toxic materials and old organelles, and recycles nutrients in eukaryotic cells. Whereas the studies on autophagy have been reported in other eukaryotic cells, its functioning in plants has not been well elucidated. We analyzed the roles of OsATG10 genes, which are autophagy-related. Two rice ATG10 genes - OsATG10a and OsATG10b - share significant sequence homology (about 75%), and were ubiquitously expressed in all organs examined here. GUS assay indicated that OsATG10b was highly expressed in the mesophyll cells and vascular tissue of younger leaves, but its level of expression decreased in older leaves. We identified T-DNA insertional mutants in that gene. Those osatg10b mutants were sensitive to treatments with high salt and methyl viologen (MV). Monodansylcadaverine-staining experiments showed that the number of autophagosomes was significantly decreased in the mutants compared with the WT. Furthermore, the amount of oxidized proteins increased in MV-treated mutant seedlings. These results demonstrate that OsATG10b plays an important role in the survival of rice cells against oxidative stresses.

• Molecules and Cells
• /
• v.39 no.10
• /
• pp.768-775
• /
• 2016

The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

• Korean Journal Plant Pathology
• /
• v.3 no.4
• /
• pp.291-298
• /
• 1987

Samples showing mosaic symptom of cowpea (Vigna sesquipedalis) with vein banding, chlorotic spot, vein yellow were collected from Chinju areas in Korea, Two viruses were distinguishable by stability in sap, host range, and relations with cells and tissues were examined under an electron microscope, Blackeye cowpea mosaic(BICMV) was sap-transmissible to 7 plant species in 2 families, Of the plants, only leguminous species were systemically infected. This virus was inactivated by heating at $50-65^{\circ}C$ for 10 min, by diluting at $10^{-4}-10^{-5}$, and aging at room temperature for 1-6 days. Preparations examined under the electron microscope by direct negative staining method(DN -method) always showed particles of flexuous filament bout 750nm in length and cytopasmic inclusions. Cytoplasmic inclusions and virus particles were also confirmed to present in the cytoplasm of a mesophyll cell by ultrathin sections of BICMV infected cowpea leaves. Cucumber mosaic virus (CMV) was transmitted by sap- inoculation on inoculated leaves of Chenopodium amaranticolor, C. quinoa producing local lesions, but non-inoculated upper leaves of Nicotiana glutinosa, Cucurbita pepo and Vigna sesquipedalis producting systemic mosaic symptoms. Electron microscopic examination of virus preparation by direct negative staining showed spherical particles of about 30nm in diameter. In ultrathin sections of CMV infected tissues, virus particles of crystalline array were found in the vacuole and a large number of virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells.

• Journal of Plant Biology
• /
• v.38 no.1
• /
• pp.79-86
• /
• 1995

Visible injury symptoms such as necrosis, chlorosis and premature senscence in the leaves of Ginkgo bloba and Pinus thunbergii treated with acid rain of pH 3.2 or below were observed. The epicuticular wax erosions were observed by SEM after exposure to acid rain of pH 2.4 and 3.2 in G. biloba and pH 4.0 below in P. thunbergii. The adaxial epidennal cells and sponge parenchyma cells were compressed, and those were distorted in the leaves of G. biloba treated with simulated acid rain of pH 3.2 or below. However, vascular tissue was intact. With increase of acidity, mesophyll cells were smaller than those of control while intercellular space in mesophyU was increased. In P. thunbergii, sponge parenchyma cells and vascular tissue except epidennis were distorted after exposure to acid rain of pH 2.4. The size change of stomata in foliar injury was not observed, but the stomatal index and size of stomatal aperture in leaves treated with acid rain increased. The stomata of injured leaf were opened in both species examined.amined.

• Journal of Plant Biotechnology
• /
• v.31 no.2
• /
• pp.145-149
• /
• 2004

We investigated the anatomical aspects of vitrification in peace poplar. Comparisons were made with regard to characteristics occurring between hyperhydric and normal shoots in shoot proliferation cultures on MS medium containing 0.2 mg/L BA. Compared with normal plants, hyperhydric plants had thick, curled, and dark green leaves. Hyperhydric stems were thicker and shorter than those of normal stems. When examined under the microscopes, the mesophyll palisade cells of hyperhydric leaves were vacuolated, whereas those of normal leaves contained normal and enriched vacuole with cytoplasm. Generally, the hyperhydric leaves showed poorly developed palisade parenchyma, and revealed irregular and bigger sized intercellular structures in both palisade and spongy parenchyma as well as epidermis cells compare to those of normal leaves. In addition, the hyperhydric leaves had lower stomatal density and bigger sized cell. Vascular tissues of hyperhydic stems were less differentiated because of poorly lignified xylem tissue. The greatly expanded cortical cells and pith appeared to be the main cause of thick stems as compared with normal stems.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.30 no.2
• /
• pp.195-200
• /
• 1985

This experiment was carried out to know the processes of protoplast isolation, culture and plant regeneration in aims of introducing foreign genes into plant cells through plant gene vector, and cellular selection for plant improvement. The main results indicated that 2% cellulase plus 0.5% macerozyme is proper for isolation of protoplasts from leaf mesophyll cells of N. plumbaginifolia, plating efficiency was higher in 1.4-2.0 x 10$^4$ cells/ml, complete cell wall was regenerated after 2 days culture, cell division and cell mass were observed after 4 days and 2 weeks, respectively, colony was developed after 3 weeks culture, addition of 1-2mg/l BA promoted shoot differentiation while root differentiation did not required hormone and seeds were harvested from more than 100 cell lines for further investigation and study.

• KSBB Journal
• /
• v.8 no.2
• /
• pp.97-103
• /
• 1993

This study was investigated as a step for the purpose of successful introduction of cytoplasmic inherited characters between the different plants. Cytoplasts were separated from the protoplasts of CMS(cytoplasmic male sterility) line such as MS Burley 21 which carried from Nicotiana megalosiphon. The cytoplasts were fused to protoplasts derived from Nicotiana tabacum Br 64 with PEG(polyethylene g1yco1). The cytoplasts were separated by density gradient centrifugation. Efficient separation of cytoplasts depended on the difference of specific density of gradient solution. However, the iso-osmolality of gradient solution was not important to separate the cytoplasts. The cells for a cybrid were fused with 50% concentration of PEG.

• Korean Journal of Environmental Agriculture
• /
• v.25 no.3
• /
• pp.268-275
• /
• 2006

ALA (5-aminolevulinic acid) has been proposed as a tetrapyrrole-dependent photodynamic herbicide by the action of the protoporphyrinogen IX oxidase (Protox IX). A study was conducted to determine photodynamic herbicidal effect of ALA on seedling growth of rice (Oryza sativa L.) and barnyard grass (Echinochloa crus-galli Beauv. var. oryzicola Ohwi) under dry and wet conditions. ALA effect on early plant growth of rice and barnyardgrass was greatly concentration dependant, suggesting that it promotes plant growth at very low concentration and inhibits at high concentration. No significant difference in herbicidal activity of biologically and synthetically produced ALAs on plant lengths of test plants was observed ALA exhibited significant photodynamic activity regardless of PSDIP and its duration. Significant shoot growth inhibition by ALA soaking treatment exhibited apparently, indicating that ALA absorbed through root system was translocated into shoot part of plants. ALA reduced plant heights of rice and barnyardgrass seedlings by 6% and 27%, respectively, showing more tolerant to ALA in rice under wet condition. Leaf thickness was reduced markedly by ALA with increasing of ALA concentration, due to mainly membrane destruction and severe loss of turgidity in mesophyll cells, although the epidermal was little affected. It was observed that photodynamic herbicidal activity of ALA applied by pre-and post-emergence application exhibited differently on plant species, and that the activity of ALA against susceptible plants was highly correlated with growing condition.

• Korean Journal of Environmental Biology
• /
• v.19 no.1
• /
• pp.1-6
• /
• 2001

Many researchers have been studied with guard cell protoplasts and detached epidermis as they think that properly stabilized protoplasts and detached epidermis retain many of the properties of intact guard cells. However, some studies have shown that stomata in detached epidermis behave differently, both quantitatively and qualitatively, from those in the intact leaf. Stomata in the intact leaf are very sensitive to environmental factors such as light, $CO_2$ and osmotic stress, but stomata in detached epidermis are less sensitive to these factors than those in the intact leaf. The clearest evidence to suggest the different response between detached epidermis and intact leaf obtained from the experiments with heavy metal, cadmium. 3-weeks old Commelina. communis was transferred to and grown in Hoagland solution in the presence or absence of 5 mM $Cd^{2+}$ for 4 days. The application of $Cd^{2+}$ showed about 70% inhibition of stomatal conductance when measured at various light intensity (100-1,000 $\mu$mole $m^{-2}s^{-1}). However, stomata in detached epidermis floated on an incubation medium containing 100 $\mu$M $Cd^{2+}$ opened to a degree of about 8.38 fm, but the stomata treated with no cadmium opened to 3.74 ${\mu}{\textrm}{m}$. These results were unexpected as the intact leaf grown in a Hoagland solution containing cadmium showed very negative physiological responses. These results showed that stomata in detached epidermis and in the intact leaf could respond reversely. Therefore, it is possible that we now misunderstand how stomata open in real natural condition.