• Title/Summary/Keyword: Explant

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Multiple shoot induction and plant regeneration from axillary buds of Magnolia 'Vulcan'

  • Kim, Tae-Dong;Kim, Ji-Ah;Lee, Na-Nyum;Choi, Chang-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.40-45
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    • 2020
  • An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia 'Vulcan' was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

Callus Induction and Plant Regeneration from Stolon in Zoysiagrass (한국잔디류에서 포복경 배양을 통한 캘러스 유기와 재분화에 관한 연구)

  • 김종보;박순정;김두환
    • Asian Journal of Turfgrass Science
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    • v.11 no.4
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    • pp.311-320
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    • 1997
  • This study was carried out to induce and maintain callus from 59 zoysiagrass lines, to know the effective disinfestation method for zoysiagrass stolon as explant and the difference in the response of callus induction among 59 lines, and to investigate the effect of medium, growth regulators, light, temperature, stolon part and internode position on callus induction and emhryogenic callus(E.C.) formation. The treatment of 0.lmg/L $HgCl_2$for 15 min resulted in no contamination and the highest callus induction(46.6%). Callus was induced from the 59 zoysiagrass lines. The callus growth of Z. japonica and Z. sinica was generally better than Z. matrella Ten cell lines whose callus and stolon grow fast in culture and in field, respectively were selected to he used for breeding. Callus induction was the most effective at 2.0mg /L of both 2, 4-D and picloram in MS medium. MS medium was the best for callus induction and growth while LS medium was the best for embryogenic callus and shoot formation. Callus induction and growth was better at 28, 31$^{\circ}C$. than 25$^{\circ}C$. and dark condition was better than light condition in MS me-dium containing 2mg/L 2,4-D. While callus induction was better with node part as explant than with internode part, callus growth and embryogenic callus formation was better with internode part. In 'Japonica 1', the first internode was the most effective in callus induction, but third internode was the best in '$M_2$ X $S_2$'.

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In vitro shoot initiation of Artocarpus heterophyllus Lam. (Jak Fruit) Effect of the explant type and the season of explant collection

  • Kahk, Kasturiarachchi;Wtpsk, Senarath;Lee, Kui-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10a
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    • pp.9-18
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    • 2003
  • A method for rapid propagation of mature Jack fruit was developed. Four types of explants (mature embryos, apical meristems of young seedlings, apices from mature plants and nodal segments) were used. It has been found 88% of young apical meristems produced shoots in Campbell and Durzan (CD) medium compared to 60% in Murashige and Skoog (MS) medium. Only 1/3 of them produced multiple shoots. Shoot idtiation from nodal segments was very rare. Mature apices produced callus. Although removed of the sheathing cover around mature buds enhanced the shoot initiation but success rate was low in growth regulator free medium. Embryos respond to the CD medium but not to the MS medium. Embryos from seeds soaked in water for 24 hours produced shoots after 8 weeks of incubation and the success rate was 70% while embryos from dry seeds only produced roots. There was no significant effect of cold storage (refrigeration) for 7 days on shoot initiation from mature embryos (65%) but the ability for shoot induction declines with storage time (55% after 21 days of cold storage). Mature axillary buds were established in Modified Campbell and Durzan (CD) medium supplemented with 0.5mg/1 and IBA. There was a significant difference in the growth performance of shoots according to the period of the year in which explants were collected. Highest (60%) was observed in November-January period. It was only 30% when the explants were collected in February-April or May-July and decreased to 20% in August-October. The shoots produced in November-January showed a higher vigor than those produced in other months. Since Jak fruit show seasonal changes in fruit bearing and shedding of leaves, it can be suggested that the difference in growth performances of tissues cultured in artificial culture media would have been affected by endogenous rhythms.

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Conceptus-derived cytokines interleukin-1β and interferon-γ induce the expression of acute phase protein serum amyloid A3 in endometrial epithelia at the time of conceptus implantation in pigs

  • Soohyung Lee;Inkyun Yoo;Yugyeong Cheon;Hakhyun Ka
    • Animal Bioscience
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    • v.36 no.3
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    • pp.441-450
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    • 2023
  • Objective: Serum amyloid A3 (SAA3), an acute phase response protein, plays important roles in opsonization, antimicrobial activity, chemotactic activity, and immunomodulation, but its expression, regulation, and function at the maternal-conceptus interface in pigs are not fully understood. Therefore, we determined the expression of SAA3 in the endometrium throughout the estrous cycle and at the maternal-conceptus interface during pregnancy. Methods: Endometrial tissues from pigs at various stages of the estrous cycle and pregnancy and with conceptuses derived from somatic cell nuclear transfer (SCNT), conceptus tissues during early pregnancy, and chorioallantoic tissues during mid- to late pregnancy were obtained and the expression of SAA3 was analyzed. The effects of the steroid hormones, interleukin-1β (IL1B), and interferon-γ (IFNG) on the expression of SAA3 were determined in endometrial explant cultures. Results: SAA3 was expressed in the endometrium during the estrous cycle and pregnancy, with the highest level on day 12 of pregnancy. The expression of SAA3 in the endometrium was significantly higher on day 12 of pregnancy than during the estrous cycle. Early-stage conceptuses and chorioallantoic tissues during mid to late pregnancy also expressed SAA3. The expression of SAA3 was primarily localized to luminal epithelial cells in the endometrium. In endometrial explant cultures, the expression of SAA3 was induced by increasing doses of IL1B and IFNG. Furthermore, the expression of SAA3 decreased significantly in the endometria of pigs carrying conceptuses derived from SCNT on day 12 of pregnancy. Conclusion: These results suggest that the expression of SAA3 in the endometrium during the implantation period increases in response to conceptus-derived IL1B and IFNG. The failure of those appropriate interactions between the implanting conceptus and the endometrium leads to dysregulation of endometrial SAA3 expression, which could result in pregnancy failure. In addition, SAA3 could be a specific endometrial epithelial marker for conceptus implantation in pigs.

Spatiotemporal expression and regulation of peptidase inhibitor 3 and secretory leukocyte protease inhibitor at the maternal-fetal interface in pigs

  • Soohyung Lee;Inkyu Yoo;Yugyeong Cheon;Hakhyun Ka
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1034-1043
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    • 2023
  • Objective: Two serine protease inhibitors, peptidase inhibitor 3 (PI3) and secretory leukocyte protease inhibitor (SLPI), play important roles in protease inhibition and antimicrobial activity, but their expression, regulation, and function at the maternal-fetal interface in pigs are not fully understood. Therefore, we determined the expression and regulation of PI3 and SLPI in the endometrium throughout the estrous cycle and at the maternal-fetal interface in pigs. Methods: Endometrial tissues during the estrous cycle and pregnancy, conceptus tissues during early pregnancy, and chorioallantoic tissues during mid to late pregnancy were obtained, and the expression of PI3 and SLPI was analyzed. The effects of the steroid hormones estradiol-17β (E2) and progesterone (P4) on the expression of PI3 and SLPI were determined in endometrial explant cultures. Results: PI3 and SLPI were expressed in the endometrium during the estrous cycle and pregnancy, with higher levels during mid to late pregnancy than during the estrous cycle and early pregnancy. Early-stage conceptuses and chorioallantoic tissues during mid to late pregnancy also expressed PI3 and SLPI. PI3 protein and SLPI mRNA were primarily localized to endometrial epithelia. In endometrial explant cultures, the expression of PI3 was induced by increasing doses of P4, and the expression of SLPI was induced by increasing doses of E2 and P4. Conclusion: These results suggest that the PI3 and SLPI expressed in the endometrium and conceptus tissues play an important role in antimicrobial activity for fetal protection against potential pathogens and in blocking protease actions to allow epitheliochorial placenta formation.

Matrix metalloproteinases: expression and regulation in the endometrium during the estrous cycle and at the maternal-conceptus interface during pregnancy in pigs

  • Inkyu Yoo;Soohyung Lee;Yugyeong Cheon;Hakhyun Ka
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1167-1179
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    • 2023
  • Objective: Matrix metalloproteinases (MMPs) are a family of endoproteases produced by various tissues and cells and play important roles in angiogenesis, tissue repair, immune response, and endometrial remodeling. However, the expression and function of MMPs in the pig endometrium during the estrous cycle and pregnancy have not been fully elucidated. Thus, we determined the expression, localization, and regulation of MMP2, MMP8, MMP9, MMP12, and MMP13 in the endometrium throughout the estrous cycle and at the maternal-conceptus interface during pregnancy in pigs. Methods: Endometrial tissues during the estrous cycle and pregnancy and conceptus and chorioallantoic tissues during pregnancy were obtained and the expression of MMPs was analyzed. The effects of steroid hormones and cytokines on the expression of MMPs were determined in endometrial explant cultures. Results: Expression levels of MMP12 and MMP13 changed during the estrous cycle, while expression of MMP2, MMP9, MMP12, and MMP13 changed during pregnancy. Expression of MMP2, MMP8, and MMP13 mRNAs was cell type-specific at the maternal-conceptus interface. Gelatin zymography showed that enzymatically active MMP2 was present in endometrial tissues. In endometrial explant cultures, estradiol-17β induced the expression of MMP8 and MMP12, progesterone decreased the expression of MMP12, interleukin-1β increased the expression of MMP2, MMP8, MMP9, and MMP13, and interferon-γ increased the expression of MMP2. Conclusion: These results suggest that MMPs expressed in response to steroids and cytokines play an important role in the establishment and maintenance of pregnancy by regulating endometrial remodeling and processing bioactive molecules in pigs.

Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars (국내 콩(Glycine max) 품종 형질전환 초기조건 확립)

  • Lee Ki-Jung;Seo Jen-Kyung;Lee Hye-Young;Jeon Eun-Hee;Shin Sang-Hyun;Lee Jai-Heon;Kim Doh-Hoon;Ko Jong-Min;Hahn Won Young;Baek In-Youl;Oh Boung-Jun;Chung Young-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.289-296
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    • 2006
  • In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.

Plant Regeneration via Organogenesis from Seed Explants in Red Pepper (Capsicum annuum L.)

  • Lee, Kwang-Woong
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.167-172
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    • 1996
  • Efficient plant regeneration has been achieved via organogenesis in the red pepper plant (Capsicum annum L.). Shoots were induced from seed explants of cultivar 'Friendship' on Murashige and Skoog's (MS) basal medium supplemetned with; NAA or IAA, and BAP or zeatin. Seed explants on the medium supplemented with 0.1-0.3 mg/L IAA and 2-5 mg/L zeatin for 2 weeks vigorously formed normal shoots in more than 90% of the explants. When these were transferred to MS medium containing 0.5-1.0 mg/L GA, 90-100% of the shoots have elongated within 1-2 weeks. The elongated shoots rooted in media supplemented with 0.3 mg/L NAA. It was revealed that this method is very rapid and efficient regeneration system for red pepper and regenerated plants can be obtained after only 5-6 weeks of culture.

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