• 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.

• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

• The Korean Journal of Mycology
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• v.24 no.3 s.78
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• pp.167-175
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• 1996

The objective of this research was to determine the effect of light quality on formation of fruit body primordia (FBPs) of Ganoderma lucidum. To achieve this 5 isolates of the fungus that develops fruit body primordia on nutrient agar media were incubated with or without continuous irradiation. The fluorescent lamps used different colors such as black light blue (BLB), pure blue (P-B), pure green (P-G), pure yellow (P-Y) and pure red (P-R). Effect of periodic light and dark exposures on FBP formation of isolate Gl-009 was investigated. The FBP formation in G. lucidum isolates was also tested under monochromatic light produced by the combination of interference filters and colored glass filters. Three isolates produced FBPs under all kinds of fluorescent lamps, whereas two induced FBPs only under visible light except for BLB fluorescent lamp. However, these isolate did not form FBPs in the dark. The FBP was formed at light intensity from 0.05 to $10.0\;{\mu}mol\;m^{-2}s^{-1}$, and begun to reduce its number as light intensity increase over $0.5\;{\mu}mol\;m^{-2}s^{-1}$. When the isolate was incubated under periodic light and dark exposures, the number and weight of FBP increased as compared with those under continuous light. Initiation of FBP requires at least 4 days of light illumination. Although isolate Gl-003 produced FBPs in a wide range of 400 to 800 nm, other four isolates had two effective regions 400 to 500 nm and 700 to 750 nm in FBP formation.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.93-97
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• 1999

Shoot tips with 2 leaf primordia were cultured to induce shoot primordia in MS liquid medium supplemented with several concentrations of BA and hIAA under the conditions of 10,000 lux illuminations for 24 h and of vertical shaking of 2 rpm in carrot. Two F$_1$ hybrids and two male sterility lines were used. Shoot primordia were only induced in the medium supplemented with 2.0 mg/L of BA and 0.2 mg/L of NAA. Genotypic specificity and seasonal effect of donor parents on shoot primordia induction were not observed and average 15-20% of the planted dornes developed to shoot primordia. The induced shoot primordia were successfully propagated by subculture in the same medium. However, they were grown into three different types during multiplication, that is, the type with multiple small shoots on the surface, the type of without any shoot, and the type of callus. Shoot primordia clusters with small shoots on the surface differentiated multiple shoots successfully in 1/2 MS solid medium supplemented with 0.2 to 1.0 mg/L of IAA and 0.2 to 1.0 mg/L of kinetin. New shoot primordia with small shoots were well formed when pieces bigger than 2 mm in diameter of the out layer of the shoot primordia cluster with small shoots were subcultured. No differences of multiplication and shooting ability and chromosomal variation of shoot primordia were observed until the 13th sub-culture.

• Journal of Forest and Environmental Science
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• v.24 no.2
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• pp.79-88
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• 2008

Twenty-eight of 44 Indian Ceropegia species are endemic and their survival is threatened. As a step towards conservation, we implied in vitro methods for the sustainable propagule production in C. hirsuta, C. lawii, C. maccannii, C. oculata and C. sahyadrica. Effects of explant, growth regulators, sucrose and photoperiod were studied. High frequency microtuber production was achieved with the seedling-apical buds, grown on MS medium containing 4-6 mg $1^{-1}$ BAP, 3-8% (w/v) sucrose, under continuous illumination. Each microtuber, when subcultured proliferated to form a cluster of secondary microtubers. Every primary and secondary microtuber bore at least one shoot-bud and a root primordium. Each tuber (formed with any of the significantly effective treatments) weighed more than 500 mg, enough to plant directly in non-sterilized soils. Microtubers could be produced and proliferated round the year. Proliferation could be solely attributed to in vitro procedures as these plants bear solitary tubers in vivo. Microtubers could be sprouted in vitro to prepare ready to pot plantlets. As, this novel method succeeded for all five species, though they belong to different eco-physiological backgrounds, we recommend its implementation in the conservation programs for a broader range of Ceropegia species, supported by other integrated strategies.

• Applied Microscopy
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• v.26 no.2
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• pp.157-175
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• 1996

The development of flexor digital tendon of the hand was studied by electron microscopy in human fetuses ranging from 9 mm to 260 mm crown rump length. The primordium of tendons was first identified as discrete collection of mesenchymal cells at 25 mm fetus. Synovial sheath formation had commenced by 40 mm fetus and was complete by 70 mm fetus. Cell junction or adhesion sites at all ages were noted between the tendon cells. When dilatation of the synovial cavity occurred, two types of synovial cells were observed. A-type cells had numerous vesicles and large vacuoles. In contrast, B-type cells were characterized by abundant rough endoplasmic reticulum and well-developed Golgi complex. By $150mm{\sim}260mm$ fetuses, a mojority of the synovial cells were type B. The most remarkable difference between the synovial cells of full-term fetus and adult was the larger amount of collagen fibers in the latter. The vascular buds were first observed between the individual fibril bundles in the interfascicular space at 150 mm fetus. At 25 mm fetus, collagen fibrils were first noted within narrow cytoplasmic recesses which were continued with the extracellular space. Collagen fibrils were filled in almost entire extracellular space at 150 mm fetus. Besides collagen fibrils in the extracellular space small elastic fibers were also identified and followed in their development.

• BMB Reports
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• v.37 no.3
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• pp.339-342
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• 2004

Germination is a process which characterized with nescient synthesis of genes. Among the genes synthesized during the germination of wheat embryos, germin genes, proteins and their enzymatic activity were defined. Germin is a water soluble homopentameric glycoprotein which is remarkable resistant to degradation by a broad range of proteases including pepsin. Germin proteins found to have strong oxalate oxidase activity which produces hydrogen peroxide by degrading oxalic acid. The current study, aimed to localize the germin genes, proteins and enzymatic activities in developing coleoptiles which is a rapidly growing protective tissue of leaf primordium and shoot apex. Non-radioactively abeled germin riboprobes were employed to localize germin mRNAs in situ. FITC (Fluorescein isothiocyanate) and alkaline phosphatase linked anti-germin antibodies were used to localize germin proteins under the fluorescence and light microscopy and finally germin enzymatic activity was localized by using appropriate enzyme assay. The results revealed that in coleoptiles germin genes, proteins and their enzymatic activity were predominantly associated with the cells of epidermis and vascular bundle sheath cells.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1055-1059
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• 2013

The ectomycorrhizal fungus Tricholoma matsutake grows symbiotically with Pinus densiflora. Phenylalanine ammonia-lyase (E.C. 4.3.1.24) catalyzes the conversion of L-phenylalanine to trans-cinnamic acid. The role of fungal phenylalanine ammonia-lyase, however, has not been clear until now. In this study, the gene encoding phenylalanine ammonia-lyase (PAL), which was isolated from T. matsutake, was cloned and characterized. The PAL gene (tmpal) consists of 2,160 nucleotides, coding for a polypeptide containing 719 amino acid residues. The deduced amino acid sequence of tmpal from T. matsutake shows high identity (70%) with that from Laccaria bicolor. Comparative analysis of the PAL genes among T. matsutake and other species of the class Agaricomycetes showed that both active sites and binding sites were significantly conserved among these genes. The transcriptional analysis of the PAL gene revealed a differential gene expression pattern depending on the developmental stages (mycelium, primordium, stipe, pileus, and gills) of T. matsutake. These results suggest that the PAL gene in T. matsutake plays an important role in multiple physiological functions.

• Animal Systematics, Evolution and Diversity
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• v.21 no.1
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• pp.21-30
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• 2005

The morphogenesis of the marine ciliate, Pelagostrobilidium simile Song and Bradbury, 1998, was investigated using pyridine silver carbonate impregnation. The morphogenesis of P. simile is of hypoapokinetal mode. The oral primordium of P. simile commences slightly below the external membranelles (EM) with the proliferation of an anarchic field. Somatic ciliature in proter and opisthe of P. simile are derived from the old structure with the proliferation of the basal bodies during the dividing process. Parental oral apparatus of P. simile is inherited by the proter, and no reorganization of oral apparatus was observed in the parental oral infraciliature.

• Journal of Mushroom
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• v.2 no.3
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• pp.145-148
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• 2004

This study was carried out to screen the fruiting body formation-specific genes from the medicinal mushroom Cordyceps militaris. A cDNA synthesized using total RNA from 4 stages of mushroom development, mycelium, primordium, immature fruiting body and mature fruiting body. Differential expression gene screening was performed by DD-PCR(Differential Display Arbitrary Primer PCR) with cDNA, we sequenced partial 6 genes using pGEM cloning vector. The DNA Sequence of the six DD-PCR products derived from differentially expressed genes was compared to that in the GenBank database by using the NCBI BLAST search to identify similarities to known sequences. Sequence analysis showed that six of DD-PCR products have unknown sequence.