• Title/Summary/Keyword: embryogenic callus induction

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The Induction of Somatic Embryogenic Callus from Petals-Derived Callus in Rosa hybrida (국내 육성 장미 품종 꽃잎 유래 체세포배 발생 캘러스 유도)

  • Lee, Su Young;Shin, Ju Young;Lee, Young Ah;Ahn, Chang Ho;Kim, Yae Jin;Park, Pil Man;An, Hye Ryun;Lee, Ka Youn;Jung, Hyun Hwan
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.652-658
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    • 2022
  • This study was conducted to induce somatic embryogenic callus (SEC) derived from petals in rose. The petal explants of 3 cultivars ('Ice Wing', 'Orange Eye' and 'Pink Beauty') with different flower colors were placed on three types media (MS, SH and WPM) supplemented with 11 mg/L 2,4-D, respectively, and then cultured in the dark for 47 days. Calluses were formed at explants of all three cultivars. Also, 'Ice Wing', which were cultured in the SH as the basal medium, showed the highest callus formation rate. However, somatic embryos were generated from only petal-derived callus of 'Ice Wing', which were induced on the WPM as the basal medium, transferred it to SH basal medium supplemented with 3 mg/L 2,4-D, and 300 mg/L L-proline, and cultured for 5 weeks. The SEC has been proliferated every four weeks at the subculture interval. In addition, as a results of making a comparison of expression of RhSERK3 and RhSERK4, which is used as signal for generation of somatic embryo from callus in rose, between the SEC and petal-derived callus from 'Ice Wing' by RT-qPCR, the former showed 10 times higher RhSERK3 expression and 700 times higher RhSERK4 expression than the latter.

Acquirement of transgenic rose plants from embryogenic calluses via Agrobacterium tumefaciens (배발생 캘러스를 이용한 아그로박테리움 매개형질전환 장미 식물체 획득)

  • Lee, Su-Young;Lee, Jung-Lim;Kim, Won-Hee;Kim, Seung-Tae;Lee, Eun-Kyung
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.511-516
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    • 2010
  • The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

Somatic Embryogenesis in Withania somnifera (L.) Dunal

  • Rani, Gita;Virk, Gurdip Singh;Nagpal, Avinash
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.113-118
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    • 2004
  • Somatic embryos were formed from calli obtained from axillary shoots (raised from nodal segments of glasshouse-grown plants under aseptic conditions), internodal segments (from in vitro-raised plants), and root and coty-ledonary leaf segments (from in vitro-raised seedlings) after 8 weeks of initial culture. Embryo formation was the highest (97.33%) from cotyledonary leaf callus on Mura-shige and Skoog's (MS) medium containing kinetin (KN) (3 mg/L). Somatic embryo induction was lesser with different combinations of auxins while it increased to 100% in internodal segment and cotyledonary leaf calli with 6-benzyladenine (BA) (2mg/L) along with 2,3,5-triiodobenzoic acid (TIBA) (2mg/L). The shoots were induced from somatic embryos raised from root, coty-ledonary leaf and internodal segment calli grown on MS medium containing BA in combination with indole-3-acetic acid (IAA). Maximum of 66.67% cultures formed shoots on MS medium containing BA (1mg/L) in combination with IAA (2mg/L). The shoots raised from somatic embryos were rooted on MS medium supplemented with indole-3-butyric acid (IBA) (2mg/L). The plantlets transferred to the field showed 70% survival rate after one year.

Development of a Protocol for Somatic Embryogenesis of Cnidium officinale M akino

  • Hui Yeong Jeong;Ji Ah Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.51-51
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    • 2021
  • This study was conducted to develop a somatic embryogenesis protocol for the Cnidium officinale Makino difficult to seed propagation. The immature flowers were used as explants. The concentration of a 2,4-D 1.0mg/L was found to be optimal concentration for induction of embryogenic callus and somatic embryos. Addition of 0.3mg/L, 0.5mg/L and 1.0mg/L to the embryo germination medium promoted somatic embryo germination. Among four concentrations, GA3 1.0mg/L were superior to others. Shoots were transferred to hormone-free MS medium after 2 months of culture in the dark. We obtained an optimized protocol for the regeneration of C. officinale.

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Plant Regeneration of Major Cultivars of Sweetpotato (Ipomoea batatas) in Korea via Somatic Embryogenesis (체세포배발생을 통한 국내 주요 고구마 품종의 식물체 재분화)

  • Kwon, Eun-Jeong;Kwon, Suk-Yoon;Kim, Moon-Za;Lee, Joon-Seol;Ahn, Young-Sup;Jeong, Byeong-Choon;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.189-192
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    • 2002
  • An efficient plant regeneration system of major cultivars of sweetpotato (Ipomoea batatas (L.) Lam.) in Korea via somatic embryogenesis was established. Embryogenic calli were formed from shoot apical meristems of sweetpotato cultivars when cultured on LS medium supplemented with 1 mg/L auxin (2,4-D, picloram, dicamba). Among three kinds of auxin, 1 mg/L 2,4-D showed the highest embryogenic calli induction rate. After 4 weeks of cultures on LS medium supplemented with 1 mg/L 2,4-D, embryogenic calli induction rates of Sinhwangmi, Zami, Yulmi, and White Star were 86%, 78%, 76%, and 80%, respectively. Upon transfer onto LS basal medium, most of somatic embryos developed into plantlets. Regenerated plantlets were transplanted to potting soil and grown to mature plants in a greenhouse.

Effect of picloram and 2,4-D on plant regeneration from mature and immature embryos of moroccan durum wheat varieties

  • Ahansal, Khadija;Aadel, Hanane;Udupa, Sripada Mahabala;Gaboun, Fatima;Abdelwahd, Rabha;Ibriz, Mohammed;Iraqi, Driss
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.131-138
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    • 2022
  • An efficient genetic transformation protocol is a fundamental requirement for high regeneration capacity from cultivated durum wheat (Triticum durum) varieties. In this study, wereportedtheeffectsoftwoauxins,2,4-dichlorophenoxyaceticacid(2,4-D)and4-amino-3,5,6-trichloropicoli nicacid(picloram), at a concentration of 2 mg/Laloneandincombination on the embryogenic callus and plantlet regeneration of four durum wheat varieties (Amria, Chaoui, Marouane, and Tomouh) using mature embryos (MEs) and immature embryos (ImEs). Significanteffectsofvariety,culturemedium(theauxinused),andvariety-mediuminteraction were observed on the callus weight and plantlet regeneration of both MR and ImE explants. The medium used for callus induction significantly affected plantlet regeneration (p < 0.001). Comparedto2,4-D, picloram led to a higher plantlet regeneration rate in both ME and ImE explants (19.8% and 40.86%, respectively). Plantlet regeneration also varied significantly depending on the variety and medium used. PicloramledtohighplantletregenerationofbothME and ImE explants in all varieties except Tomouh, which showed high plantlet regeneration of ME explants in 2,4-D. A comparison of ME and ImE responses indicated that ImEs are the best explants for high plantlet regeneration in durum wheat. Ourfindingssuggestthatpicloramisthebestauxin and should be used instead of 2,4-D due to its positive effect on increasing plant regeneration of durum wheat ME and ImE explants.

Plant Regeneration from Different Explant Types of Birdsfoot Trefoil (Lotus corniculatus L.) (버즈풋 트레포일 절편체 종류의 배양에 따른 식물체 재분화)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Kim, Meing-Jooung;Ji, Hee-Chung;Lee, Joung-Kyong;Seo, Sung;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.3
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    • pp.171-176
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    • 2008
  • Efficient plant regeneration system of birdsfoot trefoil (Lotus corniculatus L.) was development. The factors affecting the somatic embryo formation, its proliferation and regeneration capacity of leaf and stem explants of Empire cultivar was investigated. The highest number of somatic embryos was obtained on B5 medium supplemented with 1 mg/L NAA and 1 mg/L BA. Depending on different explants, highest frequency of embryogenic callus and regeneration were observed in Empire with leaf explants. The response from stem explants was slower and callus induction was less than that from leaf explants. Regenerated shoots formed complete plantlets in on 1/2 MS medium supplemented with 1 mg/L IBA. Regenerated plants were morphologically uniform with normal shape and growth pattern.

Effective Micropropagation of Pulsatilla cernua var. koreana through Apical Meristem Culture (할미꽃 정단 분열조직 배양을 통한 효율적 미세번식)

  • Ko, Jeong-Ae;Kim, Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.362-367
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    • 2008
  • In order to investigate the effect of plant growth regulators on effective in vitro micropropagation, apical meristems of Pulsatilla cernua var. koreana were cultured on Murashige and Skoog's (MS) medium with 2,4-D, NAA, TDZ and BA. Media containing 2,4-D and kinetin, 2,4-D and TDZ, NAA and TDZ were not effective on callus induction. However, embryogenic or organogenic callus was obtained on media containing NAA and BA. Especially, on MS medium with 0.5mg/L NAA and 1.0mg/L BA was optimal for a high frequency (62%) of shoot or shoot bud obtained from callus. Callus proliferation, shoot multiplication and elongation were significantly increased by adding 10% coconut water on MS media with 0.5mg/L NAA and 1.0mg/L BA. Repeated subculturing of in vitro grown shoots resulted in propagation rate of 12.9 shoots per explant every 30 days. Root formation from the adventitious shoots was not easily achieved. However, roots were only produced through callus on MS medium with 2.0mg/L NAA alone or 0.5mg/L NAA and 1.0mg/L BA. These roots were used materials for callus and shoot production repetitively.

Multi-secondary Somatic Embryogenesis and Plant Regeneration from Hypocotyl Cultures of Alfalfa (Medicago sativa L.) (알팔파의 하배축으로부터 다량의 이차 체세포배 발생과 식물체 재분화)

  • Won, S.H.;Lee, B.H.;Kim, K.Y.;Lee, H.S.;Lee, H.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.273-280
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    • 1999
  • Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

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