• Title/Summary/Keyword: explant

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Expression and regulation of prostaglandin transporters, ATP-binding cassette, subfamily C, member 1 and 9, and solute carrier organic anion transporter family, member 2A1 and 5A1 in the uterine endometrium during the estrous cycle and pregnancy in pigs

  • Jang, Hwanhee;Choi, Yohan;Yoo, Inkyu;Han, Jisoo;Kim, Minjeong;Ka, Hakhyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.5
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    • pp.643-652
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    • 2017
  • Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.

Culture and Regeneration of Populus alba × glandulosa Leaf Protoplasts Isolated from in vitro Cultured Explant (현사시나무 기내배양(器內培養) 엽육조직(葉肉組織)에서 분리(分離)된 원형질체(原形質体) 배양(培養) 및 식물체(植物体) 재분화(再分化))

  • Park, Young Goo;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.77 no.2
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    • pp.208-215
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    • 1988
  • The leaf mesophyll protoplasts of Populus alba ${\times}$ glandulosa were isolated from leaf of plantlet in vitro and cultured for plant regeneration. The MS medium (minus $NH_4NO_3$) with 0.5 mg/l BAP and 2.0 mg/l 2, 4-D showed the moderate frequency of dividing protoplasts cultured by the liquid plating method during the first week of culture. The percentage of colony formation was revealed the highest frequency by the gauze contained semi-solid agar plating method after 5 weeks cultured. Ridding out the gauze, the micro-callus was formed on the same semi-solid medium in 8 weeks after protoplasts culture. For proliferation of callus, mini-callus was transferred on the MS solid medium with 0.5 mg/l 2, 4-D and 0.1 mg/l BAP 12 weeks after culture. Shoot regeneration occurred when the calli derived from protoplasts were cultured on MS medium with 1.0 mg/l zeatin and such shoots could be readily rooted on the one half strengthen MS medium with non-phytohormone. Rooting shoots were planted in green-house 22 weeks after protoplast culture.

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Transient Expression of β-gulucuronidase (GUS) gene in Immature Ovules and Calli Derived from Cottonwood Species (Populus deltoides) by Microprojectile Bombardment (포플러의 미성숙(未成熟) 배(胚)와 캘러스에서 유전자총(遺傳子銃)에 의(依)한 GUS-gene의 일시적(一時的) 발현(發現))

  • Kang, Hoduck;Kang, Sang-Gu;Bae, Hanhong;Park, Kyo-Soo;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.86 no.3
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    • pp.261-269
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    • 1997
  • Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and ${\beta}$-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced about $25.75{\pm}2.77$(per ovule), $11.43{\pm}1.22$(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.

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Cyclosporin A-induced Gingival Overgrowth is Closely Associated with Regulation Collagen Synthesis by the Beta Subunit of Prolyl 4-hydroxylase and Collagen Degradation by Testican 1-mediated Matrix Metalloproteinase-2 Expression

  • Park, Seong-Hee;Kim, Jae-Yoen;Kim, Hyun-Jeong;Park, Kwang-Kyun;Cho, Kyoo-Sung;Choi, Seong-Ho;Chung, Won-Yoon
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.205-211
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    • 2008
  • Gingival overgrowth can cause dental occlusion and seriously interfere with mastication, speech, and dental hygiene. It is observed in 25 to 81% of renal transplant patients treated with cyclosporine A (CsA). CsA-induced gingival overgrowth (CIGO) is caused by quantitative alteration of the extracellular matrix components, particularly collagen. However, the molecular mechanisms involved in the pathogenesis of CIGO remain poorly understood, despite intense clinical and laboratory investigations. The aim of the present work is to identify differentially expressed genes closely associated with CIGO. Human gingival fibroblasts were isolated by primary explant culture of gingival tissues from five healthy subjects (HGFs) and two patients with the CIGO (CIGO-HGFs). The proliferative activity of CsA-treated HGFs and CIGO-HGFs was examined using the MTT assay. The identification of differentially expressed genes in CsA-treated CIGO-HGF was performed by differential display reverse transcriptase-polymerase chain reaction (RT-PCR) followed by DNA sequencing. CsA significantly increased the proliferation of two HGFs and two CIGO-HGFs, whereas three HGFs were not affected. Seven genes, including the beta subunit of prolyl 4-hydroxylase (P4HB) and testican 1, were upregulated by CsA in a highly proliferative CIGO-HGF. The increased P4HB and testican-1 mRNA levels were confirmed in CsA-treated CIGO-HGFs by semiquantitative RT-PCR. Furthermore, CsA increased type I collagen mRNA levels and suppressed MMP-2 mRNA levels, which are regulated by P4HB and testican-1, respectively. These results suggest that CsA may induce gingival overgrowth through the upregulation of P4HB and testican-1, resulting in the accumulation of extracellular matrix components.

Chromosome Doubling of Allium wakegi Araki by In Vitro Cultures (기내배양에 의한 쪽파의 체세포 염색체 배가)

  • Yim, Sun-Hee;Ahn, Chang-Soon;Jeong, Chang-Nam;Han, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.259-264
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    • 2002
  • Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n+4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.

Micropropagation of Juvenile and Mature Tree of Corylopsis coreana by Axillary Bud Culture (액아배양에 의한 유묘 및 성숙 히어리나무의 기내번식)

  • Moon, Heung-Kyu;Noh, Eun-Woon;Ha, Yoo-Mi;Shim, Kyung-Ku
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.117-121
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    • 2002
  • We have developed an in vitro micropropagation system via shoot formation from axillary buds using nodal segments of Corylopsis coreana. Explants from both juvenile tree (one-year-old greenhouse stock seedlings) and mature tree (ten-years-old tree in nursery) were compared with regard to propagation efficiency. Combined treatment of both BA and zeatin were effective on shoot proliferation since the best result was obtained on MS medium supplemented with 0.5∼3.0 mg/L zeatin and 0.2 mg/L BA. Generally, juvenile explants were better in both shoot proliferation and growth than mature explants. However, as the duration of in vitro culture was proceed to 6 months, explants from mature tree also produced three shoots per explant. Distinctive differences in rooting and adaptability to soil of shoots obtained from mother trees. Whereas shoots originated from juvenile explants rooted as high as 97%, those from adult explants showed 62% rooting. Similar result was also observed in soil acclimatization. The plantlets derived from juvenile plants survived 67%, while only 48% of those from adult trees survived. The results showed a possibility of the micropropagation of Corylopsis coreana through shoot formation from axillary buds. In addition, the advance of the research still remain to enhance the frequency of acclimatization of plantlets from mature trees for practical application.

Endoreduplication Pattern of Somatic Embryos and Variants Occurrence Affected by Pre-existed Endoreduplicated Cells in Doritaenopsis (Doritaenopsis 체세포배의 내배수성 특성과 절편체의 내배수성 세포에 기인한 체세포변이의 발생)

  • Park, So-Young;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.297-302
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    • 2006
  • In general, the proliferation of orchids via somatic embryos has been used for mass production of somatic clones because of high propagation efficiency. In spite of high propagation rate, this method often brings somaclonal variation, especially polyploid frequency. Therefor we here concentrated to investigate the relationship between endopolyploidization patterns of explants and the occurrence of tetraploid variant in clonally proliferated Doritaenopsis via somatic embryo regeneration system. In the fully developed somatic embryo, upper part contained 2C to 16C while middle and lower parts showed 2C to 32C DNA content. Two-week-old embryo contained 2C to 16C, whereas those regenerated after 4 to 10-week-old contained 2C to 64C nuclei. Results showed that endoreduplication was variable depending upon tissue types, ages, and parts in one species. lower part of somatic embryo having high endoreduplication degree increased the regeneration of tetraploid variants by about 3-fold comparing to upper part of somatic embryo culture. polyploid frequency occurrence might be closely related to the high levels of endoreduplication of somatic embryos used as explant. It suggested that the upper part of somatic embryo having comparatively low endoreduplication degree is suitable for the stable in vitro propagation system.

In vitro shoot propagation of Ranunculus kazusensis Makino, an endangered aquatic plant (멸종위기 수생식물인 매화마름(Ranunculus kazusensis Makino)의 기내 신초 증식)

  • Park, Min Wan;Ryu, Shi Hyun;Nam, Su Hwan;Bae, Kee Hwa
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.325-329
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    • 2017
  • A micropropagation method via callus for Ranunculus kazusensis Makino, an endangered species, was established. When stem segments were cultured on MS media supplemented with 1.0 mg/L IAA, NAA, IBA and 2,4-D, the highest frequency of callus induction was achieved on MS medium supplemented with 1.0 mg/L NAA. Multiple shoot per explant was obtained, the MS medium containing 1.0 mg/L BA and 0.5 mg/L NAA. Additionally, effect of activated charcoal (AC) and sucrose on shoot growth in in vitro culture were examined. The most suitable conditions for shoot growth after 4 weeks of culture were the MS medium with AC and sucrose. This in vitro propagation protocol will be valuable for conservation and mass propagation of this endangered plant.

Plant Regeneration via Adventitious Shoot Formation in Platycodon grandiflorum (Jacq. A. DC.) (도라지 (Platycodon grandiflorum (Jacq.) A. DC.) 부정아 형성을 통한 식물체 재분화)

  • Kim, Ju Young;Na, Hyun Sun;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.330-334
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    • 2017
  • To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog's (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1/2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).

Efficient Callus Induction and Plant Regeneration of Guineagrass (Panicum maximum Jacq.) (난지형 목초 기니아그라스의 효율적인 캘러스 유도 및 식물체 재분화)

  • Seo, Mi-Suk;Takahara, Manabu;Takamizo, Tadashi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.30 no.4
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    • pp.283-290
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    • 2010
  • Guineagrass (Panicum maximum Jacq.) is an important warm-season forage grass as well as biomass crop. It has both sexual and asexual mode of reproduction (apomictic) depending on cultivar. We developed efficient plant regeneration system for an apomictic (cv. Natsukaze) and a non-apomictic (Noh-PL1) guineagrass by optimizing the level of L-proline in the callus induction and that of $AgNO_3$ in plant regeneration medium. Among the L-proline concentrations tested, the best callus induction was achieved by using 2g/L L-proline in both the genotypes. Immature embryos proved to be the best explant source for tissue culture of guineagrass. The highest frequency of shoot regeneration was obtained on MS plant regeneration medium supplemented with 2 mg/L $AgNO_3$. These results provide a foundation for efficient tissue culture and genetic improvement of guineagrass.