• Title/Summary/Keyword: endosperm cell

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Micropropagation of Medicinal Woody Eleutherococcus pedunculus via Somatic Embryogenesis

  • Choi, Yong Eui
    • Journal of Forest and Environmental Science
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    • v.23 no.1
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    • pp.5-9
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    • 2007
  • Zygotic embryos just after harvest of seeds were immature globular to heart stage. Maturation of zygotic embryos rapidly proceed when zygotic embryos together with small excised parts of endosperm were cultured on 1/3-strength MS solid medium with 2% sucrose, and the zygotic embryos were germinated within two months. Embryogenic callus was formed from the excised segments of germinating zygotic embryos of Eleutherococcus pedunclus on Murashige and Skoog (MS) medium with $4.5{\mu}M$ 2,4-D. The embryogenic callus formation occurred at a low frequency (less than 7%) from hypocotyl segments. The embryogenic calli were maintained on the same medium as primary medium. High frequency somatic embryogenesis was obtained after the cells were transferred to medium lacking 2,4-D. Cotyledonary embryos were germinated and converted into plantlets on medium with $20{\mu}M$ $GA_3$. Embryogenic callus and somatic embryos were produced spontaneously on the surfaces of roots and/or hypocotyls of plantlets. The frequency of embryogenic callus formation was 85% in roots and 34% in hypocotyls. Therefore maintain of cell lines performed very easily. Plantlets with developed epicotyls at more than 3 cm acclimatized at high frequency (89%). While plantlets with small epicotyls (less than 1 cm) were acclimatized at low rate (32%). The soil survived plantlets produced new sprouts after over wintering in the field.

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Production of Artificial Seeds by Alginate-encapsulation of Rice Somatic Embryos (벼의 수화겔 인공종자 생산)

  • 정원중;민성란;송남희;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.183-186
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    • 1994
  • Somatic embryos derived from cell suspension cultures of rice were singly alginate-encapsulated to be used as artificial seeds. When placed on half strength MS solid medium,73% of the encapsulated somatic embryos were capable of germination Encapsulation per se did not affect the germination frequency of embryos. When incubated by wrapping with moistured non-sterile filter paper, 60% of the encapsulated somatic embryos germinated. However encapsulated zygotic embryos without endosperm showed a high germination frequency regardless of the sterility of the incubation conditions. The results suggest that a greater susceptibility of somatic embryos to contaminants is attributed to lower germination frequency of encapsulated somatic embryos in non-sterile conditions.

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Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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Effects of NSP Degrading Enzyme on In vitro Digestion of Barley

  • Li, W.F.;Sun, J.Y.;Xu, Z.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.122-126
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    • 2004
  • A digestion trial in vitro was conducted to study effects of supplementation of NSP (non-starch polysaccharides) degrading enzyme (feed grade) on cell wall degradation and digestibility of nutrients in barley. The slices of barley were soaked in distilled water with or without 0.15% non-starch polysaccharides degrading enzyme. Microscopic examination of the slices showed that the endosperm cell wall of barley was completely degraded by the non-starch polysaccharides degrading enzyme. The residues and supernatant of digesta in vitro were separated by filtration with 0.1 mm nylon fabric. The residues were used for measurement of crude protein, crude fat, crude fiber, and moisture. The supernatant was used for determination of viscosity, as well as amino-nitrogen and glucose content. The results showed that compared with the control, the amino-nitrogen and glucose content of the supernatant increased by 17.58% (p<0.05) and 10.26% (p<0.05), respectively, while viscosity did not change. Enzyme supplementation increased the digestibilities of dry matter, crude protein, nitrogen-free extract, crude fat and crude fiber of barley by 18.1% (p<0.05), 20.3% (p<0.05), 16.4% (p<0.05), 26.9% (p<0.05) and 30.0% (p<0.05), respectively. The present study suggests that cell wall hydrolysis may contribute to improved nutrient digestion in vivo when non-starch polysaccharides degrading enzymes are fed to swine.

Effect of Red Light on Changes of Embryo Tissue of Barley during Germination (맥아제조시(麥芽製造時) 적색광(赤色光) 희사(熙射)에 의한 배유조직(胚乳組織)의 변화(變化))

  • Kim, Jin-Gu;Shin, Seung-Lyeul;Kim, Ju-Nam;Kim, Soon-Dong;Kim, Kwang-Soo
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.60-64
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    • 1987
  • This study was carried out to investigate the effect of the red light on embryo tissue of barley during germination. The solubility of starch in endosperm of germinated barley was different between dark and red treatments at the 3rd day of germination, but was increased by the red light from the 4th day of germination. Blue value of the starch in the germinated barley decreased rapidly from 0.42 at the 1st day to 0.13 at the 6th day in the dark, and same tendency was found in the red light, but blue value was lower in the red light than in the dark. Aleurone cell wall was swollen much faster in the red light than in the dark during germination. The cell wall was broken down more greatly in the red light than in the dark at the 5th day of germination.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • 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.

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Physicochemical and Gelatinization Properties of Glutinous Rice Flour and Starch Steeped at Different Conditions (수침한 찹쌀가루와 전분의 이화학적 및 호화 특성)

  • 최은정;김향숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.17-24
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    • 1997
  • The effects of steeping on the physicochemical and gelatinization characteristics of glutinous rice flour and its starch were studied. Steeping conditions were 1 day at 25"C,7 days at 2iC and 7 days at 35"C. Crude protein, lipid and ash content were decreased br steeping. It was observed with scanning electron microscopy that endosperm cell wall of glutinous rice flour was diminished by steeping. Although morphology of the glutinous rice starch granules was not affected, the size was decreased by steeping. Density and water binding capacity(WBC) of glutinous rice flour and its starch were changed by steeping. X-ray diffraction pattern of glutinous rice starch was A type and was not affected by steeping. Swelling power of glutinous rice flour and its starch was increased but solubility was decreased by steeping. In Brabender amylographic examination, peak viscosity of untreated glutinous rice flour was very low and increased enormously by steeping resulting in the similar Brabender viscosity pattern to its starch. The gelatinization temperature examined by X-ray diffractometry was lowered by steeping. And the degree of gelatinization under the conclusion temperature increased with increasing of steeping Period and temperature.mperature.

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Genotypic Variations in ${\beta}-glucan$ Content of Barley Cultivated in Different Regions

  • Kim Hong-Sik;Park Kwang-Geun;Baek Seong-Bum;Nam Jung-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.4
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    • pp.335-339
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    • 2006
  • The level of ${\beta}-glucan$ which is a major soluble dietary fiber found in the grain endosperm cell wall was highly variable among 25 barley genotypes grown at four locations including Suwon, Naju, Jinju, and Jeju. Statistically significant genotypic effects were observed for ${\beta}-glucan$ content at each or across growing sites (P<0.001). On average, 'Chalssalbori' showed the lowest percentage ${\beta}-glucan$ (4.04%) among genotypes in the grain, whereas 'Yonezawa Mochi' was highest in percentage ${\beta}-glucan$ (6.46%) compared to other genotypes. The significant difference between genotypes was approximately 1-2% across environments. The effects of location or interaction between locations and genotypes were not significant on the variation of ${\beta}-glucan$ contents. High ${\beta}-glucan$ content seemed to be greatly associated with such grain traits as waxiness and presence of husk except for 'Chalssalbori'. The waxy genotypes had a mean of 5.37% and values ranging from 5.28 to 5.47%, but normal genotypes had a mean of 4.78% and values ranging from 4.69 to 4.88% over environments. Hulless barley genotypes were also higher than hulled barley genotypes for the average ${\beta}-glucan$ content in both individual and over all environments. The difference between the hulled and hulless gene pools was on average of 0.37% with ranges from 0.19% to 0.56% at four environments. ${\beta}-glucan$ content measured from a mapping population of $F_5$-derived 107 lines derived from the cross between 'Yonezawa Mochi' and 'Neulssalbori' was not significantly associated with other agronomic traits except for 1,000-kernel weight at the '01 Suwon environment. Not too much information on the relationship of ${\beta}-glucan$ content to agronomic traits was available.

Taxol Production in Taxus sap. Cell Cultures 1. Studies on Taxol Content in Yew Trees and Cultured Plant Cells (주목 세포배양에 의한 Taxoll 생산 1.주목 부위 및 서식 고도별 Taxol 함량 및 세포주 유도에 관한 연구)

  • 강인선;전정욱
    • KSBB Journal
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    • v.9 no.3
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    • pp.299-305
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    • 1994
  • Taxol contents in various parts of 15 years old yew tree were determined. The descending order of taxol content per unit mass was stem bark, root bark, needle and seed. In the seed, that order was seed coat, embryo and endosperm. The total amount of taxol extractable from a 15 years old yew tree was 1.68 gram. This amount was distributed in needle, stem bark, root bark and seeds as 48.0, 23.8, 27.9 and 0.4%, respectively. Altitudinal variation of taxol content was also observed. More taxol was observed in yew trees grown at high altitude over 1000m above sea level. Calli and suspension cultures were induced from various yew trees. The presence of taxo] in cultured cells was established bv HPLC. The taxol content in cultured cells were different according to the source of explants. These results may be useful for the goal of large scale taxol production by cultured yew tree cells.

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Sturcture of the Rice Glutelin Multigene Family and Its Expression (쌀 Glutelin 유전자군의 구조 및 발현조절)

  • 황영수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.261-282
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    • 1987
  • Plants store a significant amount of their nitrogen, sulfur and carbon reserves as storage proteins in seed tissues. The major proteins present in rice seeds are the glutelins. Glutelins are initially synthesized at 4-6 days postanthesis and deposited into protein bodies via Golgi apparatus. Based on nucleic acid sequences and Southern blot analysis, the three isolated glutelin genomic clones were representative members of three gene subfamilies each containing 5 to 8 copies. A comparison of DNA sequences displayed by relevant regions of these genomic clones showed that two subfamilies, represented by clones, Gt1 and Gt2, were closely, related and probably evolved by more recent gene duplication events. The 5' flanking and coding sequences of Gt1 and Gt2 displayed at least 87% homolgy. In contrast, Gt3 showed little or no homolgy in the 5' flanking sequences upstream of the putative CAAT boxes and exhibited significant divergence in all other portions of the gene. Conserved sequences in the 5' flanking regions of these genes were identified and discussed in light of their potential regulatory role. The derived primary sequences of all three glutelin genomic clones showed significant homology to the legume 11S storage proteins indicating a common gene origin. A comparison of the derived glutelin primary sequences showed that mutations were clustered in three peptide regions. One peptide region corresponded to the highly rautable hypervariable region of legume peptide region of legume 11S storage proteins, a potential target area for protein modification. Expression studies indicated that glutelin mRNA transcripts are differentially accumulated during endosperm development. Promoterss of Gt2 and Gt3 were functional as they direct transient expression of chloramphenicol acetyltransferase in cultured plant cell.

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