scholarly journals DIRECT SOMATIC EMBRYOGENESIS AND PLANT REGENERATION FROM IMMATURE OVULES IN YOOZA(CITRUS JUNOS SIEB. ET TANAKA)

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 694c-694
Author(s):  
Sung-Do Oh ◽  
Won-Seob Song ◽  
Man-Sang Lee

From one week through 7 weeks after artificial pollination, immature ovules of yooza(Citrus junos Sieb. et Tanaka) were excised and cultured in vitro on MT media. Even though there was only a little difference in percentage of somatic embryo formation depending upon the time of excision, immature ovules of 4-week-old showed the highest ratio of somatic embryo formation without callus outgrowth. Various growth regulators or other stimulators were added to the MT media to increase the somatic embryogenesis, In general, BAP was more effective than 2,4-D for somatic embryo formation and the combinations of 0.01mg/l 2,4-D and 0,01 or 0.1mg/l BAP were particularly effective in stimulating somatic embryo formation. When 500mg/l malt extract was added to the medium, the percentage of somatic embryo formation increased reaching as high as 86.7%. Plant regeneration from somatic embryos reached to 66.7% on the medium containing 1.0mg/l zeatin. Isozyme banding patterns were also analyzed to confirm the variations of characteristics of the plantlets derived from direct somatic embryos.

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.


Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.


1992 ◽  
Vol 70 (6) ◽  
pp. 1186-1192 ◽  
Author(s):  
R. Gill ◽  
Praveen K. Saxena

An efficient procedure has been developed for inducing direct somatic embryogenesis, organogenesis, and regeneration of plants from tissue cultures of peanut (Arachis hypogaea L.). Thin transverse sections of the cotyledons and juvenile leaves were cultured on Murashige and Skoog medium supplemented with N6-benzylaminopurine (BAP) or a substituted phenylurea, thidiazuron (TDZ). Somatic embryos or shoot buds differentiated from cut surfaces of the cotyledons and midrib region of the leaves. The application of BAP induced differentiation of shoot buds whereas the treatment with TDZ resulted in the production of somatic embryos. Somatic embryos developed into plants after subculturing on a basal meduim. Agar-solidified medium was found to be superior to the liquid medium for the development of embryos and shoot buds. The procedure of TDZ-induced somatic embryogenesis and plant regeneration was successfully applied to three genotypes of peanut. A distinct feature of this study is the induction of the morphogenic competence in cultures of seedling expiants of peanut that so far have remained recalcitrant to somatic embryogenesis in vitro. Key words: peanut, Arachis hypogaea, shoot regeneration, somatic embryogenesis, thidiazuron, plant regeneration.


Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 38 ◽  
Author(s):  
Ghulam Raza ◽  
Mohan B. Singh ◽  
Prem L. Bhalla

The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration. Overall, nine soybean cultivars belonging to different maturity groups were evaluated. Immature cotyledon ranging from 2–4 and 4–6 mm in size were used as explants for somatic embryogenesis induction. Maximum somatic embryo induction frequency (86%) was observed from 4–6 mm immature cotyledons of the cv. Jack (MG III), followed by 66%, 26%, 21%, and 6% in cultivars Williams (MG III), Snowy (MG III), MoonB1 (MG V), and PNR791 (MG V), respectively. On the other hand, cv. Snowy showed maximum somatic-embryo-inducing potential (67%) in 2–4 mm immature cotyledons followed by Williams, Jack, MoonB1, and PNR791. Somatic embryos from Jack, Williams, and Snowy cultivars were further tested for embryo proliferation, maturation, and germination. Maximum proliferation and maturation were observed in cv. Jack, followed by Snowy and Williams. However, cv. Snowy showed a significantly higher conversion of cotyledonary stage embryos to plantlets (85%), than both Jack and Williams cultivars (53% each). In conclusion, this study outlined a protocol for somatic embryogenesis and plant regeneration from three soybean cultivars. Our findings suggest commercial cv. Snowy could be a good candidate for developing transgenic plants through somatic embryogenesis.


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1066D-1067
Author(s):  
Jae-Dong Chung ◽  
Hong-Yul Kim ◽  
Jung-Hae Suh ◽  
Oh-Chang Kwon ◽  
Chang-kil Kim

Somatic embryo formation was observed on thin-sectioned leaf explants within 3 weeks of culture from two Phalaenopsis hybrids—Phalaenopsis Hwafeng Redjewell `Ching Ruey' Phalaenopsis Chingruey's Giant Ching Ruey' (R×R), and Phalaenopsis Formosa Best Girl Ching Ruey' Depts. Lih Jiang Beauty `S 566' (WR×WR). Frequency of somatic embryo formation was higher in hybrid WRxWR than R×R and optimal concentration of TDZ for the induction of somatic embryos was 9.08 μM. In (WR×WR) embryo proliferation was simultaneously observed after transferring the explants with somatic embryo clumps onto PGR-free half-strength MS medium. Six months after initiation, the culture plantlets were produced. This is the first report on somatic embryogenesis induced directly from the leaf explants using TDZ in Phalaenopsis.


2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>


2016 ◽  
Vol 3 (2) ◽  
pp. 71
Author(s):  
Nur Ajijah ◽  
RR. Sri Hartati

<p><em>Information on the effect of cytokinins on cacao (</em>Theobroma cacao<em> L.) primary somatic embryogenesis and its interaction with explant types and genotypes is not yet known. This study aimed to evaluate the effect of cytokinins and its interaction with explant types and genotypes on cacao somatic embryogenesis. The study was conducted at tissue culture laboratory of IAARD, Bogor from April until December 2012 and October 2014 until February 2016. Three types of cytokinins i.e. kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M), thidiazuron (0.01, 0.02, and 0.04 </em><em>μ</em><em>M) and benzylaminopurine (0.55, 1.11, and 2.22 </em><em>μ</em><em>M) in combination with 9 </em><em>μ</em><em>M 2,4-D were tested for their effectiveness in inducing somatic embryogenesis from petals and staminoid explants of Cimanggu 1 genotype. Furthermore, three levels of kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M</em><em>) also in combination with 9 </em><em>μ</em><em>M 2,4-D were evaluated for their influences on the somatic embryogenesis from petals and staminoid explants of three cacao genotypes i.e. Sulawesi 02, ICCRI 04 and Cimanggu 3. The result demonstrated that 2.32 </em><em>μ</em><em>M kinetin and staminoids explant were more effective to induce cacao somatic embryogenesis of Cimanggu 1 genotype (7%, 0.23 embryos/explant). Additionally, there were interaction effects between the level of kinetin with explant types and genotype on the percentage of explants forming embryo at 12 weeks after culture. The highest percentage of somatic embryo formation was shown by ICCRI 04 genotype with the use of petals explant and a kinetin level of 1.16 </em><em>μ</em><em>M (31.85%), but not significantly different from the level of kinetin 2.23 </em><em>μ</em><em>M (25.55%). The formation of primary somatic embryos of cacao is largely determined by the type and level of cytokinins, type of explant, and genotype.</em></p>


1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006


2017 ◽  
Vol 17 (2) ◽  
pp. 91
Author(s):  
Meutia Zahara

Abstract Phalaenopsis orchids are recognized as the most popular orchid genus in the world, especially in horticultural industry due to their large, colorful, and durable flowers as well as their wider adaptability to room conditions. The characteristics of seedling propagated by vegetative means are not uniform; therefore, propagation through tissue culture is desirable. Although the micro propagation of Phalaenopsis has shown very good development, but the wide spread of micro propagation still limited due some problems such as the exudation of phenolic compounds, the PGR concentration, the media used, somaclonal variation, the chosen explants, etc. This paper endeavor to include some important investigations based on the common explants used; leaf and flower stalk. Keywords: Micropropagation, Phalaenopsis, leaf explant, flower stalk ReferencesAnonymous. Orchid (Orchidaceae). Diakes tanggal 13 Januari 2013 dari http://www.rainforest-alliance.org/kids/species-profiles/orchid. 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S.;Kakuta, S.; Kano, A.; Okabe, M.Efficient propagation of protocorm-like bodies of Phalaenopsis in liquid medium. Plant Cell, Tissue and Organ Culture. 1996, 45, 79–85.Park, S. Y. ; Yeung, E. C.; Chakrabarty, D. ; Paek, K. Y. An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture. Plant Cell Reports. 2002, 21, 46–51.Zahara, M.; Datta, A.; Boonkorkaew, P. Effects of sucrose, carrot juice and culture media on growth and net CO2 exchange rate in Phalaenopsis hybrid ‘Pink’. ScientiaHorticulturae. 2016,205, 17–24.Hee, K. H.; Loh, C. S.; Yeoh, H. H. In vitro flowering and rapid in vitro embryo production in Dendrobium Chao Praya Smile (Orchidaceae). Plant Cell Reports. 2007, 26, 2055–2062.Kannan, N. An in vitro study on micropropagation of Cymbidium orchids. Current Biotica. 2009, 3, 244–250.Steward, Jr. N. C. Plant Biotechnology and Genetics. Willey, A john Willey & Sons, INC., Publication. 2008.George, E. F.; Sherington, P. D.Biotechnology by tissue culture. Exegetics Ltd. 1994.Nursyamsi. Teknik kultur jaringan sebagai alternatif perbanyakan tanaman untuk mendukung rehabilitasi lahan. Makalah pada ekspose hasil-hasil penelitian balai penelitian kehutanan makasar. Makasar, 2010.Aditi, J. F. L. S.; Krikorian, A. D. Orchid mircropropagation: the path from laboratory to commercialization and an account of several unappreciated investigators. Botanical Journal of of the Linnean Society. 1996, 122: 183-241.Gunawan, L. W. Teknik Kultur Jaringan Tanaman. Pusat Antar Universitas (PAU) Bioteknologi IPB. 1998. Bogor.Chugh, S. Guha, S.; Rao, I. U. Micropropagation of orchids: A review on the potential of different explants. Scientia Horticulturae. 2009, 122, 507–520.Ramdan. Kultur daun dan pangkal batang in vitro anggrek bulan raksasa (Phalaenopsis gigantea J.J.Smith) pada beberapa media kultur jaringan. Departemen agronomi dan hortikultura, Fakultas pertanian IPB. 2011.Latip, M. A. R.; Murdad, Z. A.; Aziz, L. H.; Ting, L. M.; Govindasamy.; R. Pipin. Effects of N6-Benzyladenine and Thidiazuron on Poliferation of Phalaenopsis gigantea Protocorm. AsPac J. Mol. Biol. Biotechnol. 2010, 18(1): 217-220 p.Niknejad, A.; Kadir, M. A.; Kadzimin, B. S. In vitro plant regeneration from protocorms-like bodies (PLBs) and callus of Phalaenopsis gigantea (Epidendroidaceae: Orchidaceae). African Journal of Biotechnology.2010, 10, 11808–11816.Chen, J. T.; Chang, W. C. Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biologia Plantarum. 2006, 50, 169–173.Zahara, M. Disertasi doktor: The Effects of Plant Growth Regulators and Natural Additives on Direct Shoot Regeneration and Plantlet Growth of Phalaenopsis hybrid ‘Pink’. Asian Institute of Technology, Pathumthani. Thailand. 2016.Xu, C. J.; Li, H.; Zhang, M. G. Preliminary studies on the elements of browning and the changes in cellular texture of leaf explant browning in Phalaenopsis. Acta Horticulturae Sinica. 2005, 32, 1111–1113.Tokuhara, K; Mii, M. Induction of embryonic callus and cell suspension culture from shoot tips excised from flower stalk buds of Phalaenopsis (Orchidaceae). In Vitro Cellular & Developmental Biology–Plant. 2001, 37, 457–461Balilashaki, K.; Naderi, R.; Kalantari, S.; Soorni, A. Mircropropagation of Phalaenopsis amabilis cv Cool ‘Breeze’ with using flower stakl nodes and leaves of sterile obtained from node cultures. IJFAS, 2014.Semiarti, E.; Indrianto, A.; Purwanto, A. Agrobacterium-Mediated transformation of Indonesian orchids for  micropropagation, genetic transformation, Prof. MarÃa Alvarez (Ed.), ISBN: 978-953-307-364-4, InTech, 2011. Available from: http://www.intechopen.com/books/ genetic-transformation/agrobacterium-mediated-transformation-ofindonesian-orchids-for-micropropagation.


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