scholarly journals Effects of Osmolytic Agents on Somatic Embryogenesis of Saffron (Crocus sativus L.)

2015 ◽  
Vol 7 (1) ◽  
pp. 57-61 ◽  
Author(s):  
Maryam VAHEDI ◽  
Siamak KALANTARI ◽  
Seyed ALIREZA SALAMI

A protocol for callus induction from meristem tissues and subsequent somatic embryo formation were established in this study. Explants were taken from apical and lateral meristems of saffron and these explants were cultured on MS medium supplemented with combinations of 2.4-dichlorophenoxyacetic acid (2.4-D) and Kinetin (Kn). The effects of osmotic agents such as abscisic acid (ABA), polyethylene glycol (PEG) and Gelrite on somatic embryogenesis were also investigated. After 45 and 60 days of culture, calli were induced from apical and lateral meristems, respectively. The apical meristems yielded higher quality calli when compared to the lateral meristems. The highest frequency of callogenesis and the growth rate of callus were achieved from apical meristems on Murashige and Skoogs (MS) medium supplemented with 2.4-D (2 mg/l) and Kinetin (0.5 mg/l). After 45 days of subculture, the segments of nodular calli were transferred to plant growth regulator (PGR)- free media for induction of pre-embryogenesis embryo formation. Pre-matured embryos were cultured on MS medium supplemented with different osmotic agents such as Gelrite, ABA and PEG to study their effects on embryo maturation. Both PEG and ABA proved more effective for somatic embryo maturation as compared to Gelrite.

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.


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


1989 ◽  
Vol 19 (2) ◽  
pp. 285-288 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko

Tissue cultures were initiated from developing seeds of black locust (Robiniapseudoacacia L.) collected from three trees at weekly intervals from 1 week following anthesis until early fruit maturity. Explants were cultured on media containing 0, 2, or 4 mg/L 2,4-dichlorophenoxyacetic acid and 0 or 0.25 mg/L 6-benzyladenine. Seeds explanted onto hormone-supplemented media remained on these media for 1 or 3 weeks before being placed on hormone-free media, or were maintained on hormone-supplemented media for the entire study. Direct somatic embryogenesis was observed in a single culture, initiated from a seed collected 4 weeks after anthesis and cultured for 1 week on a medium supplemented with 4 mg/L 2,4-dichlorophenoxyacetic acid and 0.25 mg/L 6-benzyladenine before transfer to basal medium. Although it could not be discerned from which part of the explant somatic embryos were derived, secondary embryogenesis continued from the radicles of cotyledonary-stage somatic embryos. Most somatic embryos were well formed, with two distinct cotyledons. Embryos germinated precociously, producing plantlets that were initially weak but later gained vigor and resembled seedlings.


1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.


Forests ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1335
Author(s):  
Shuaifei Jiang ◽  
Xiaoyi Chen ◽  
Ying Gao ◽  
Ying Cui ◽  
Lisheng Kong ◽  
...  

Prince Rupprecht’s larch (Larix principis-rupprechtii Mayr) is a native conifer in North China with great economic and ecological values. Somatic embryogenesis (SE) is a powerful tool for the mass clonal propagation in plants. In this study, we described a high-efficiency SE system via indirect pathways and investigated the effect of genotype, culture conditions and phytohormones on SE. Immature zygotic embryos (IZEs) of L. principis-rupprechtii Mayr were used as explant materials. In the induction stage, embryogenic tissues (ETs) were induced on mLV medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzylaminopurine (6-BA). The initiation frequencies showed significant differences (p < 0.05) among 20 genotypes of open-pollinated mother trees with the highest induction frequency reaching 30%. For tissue proliferation, proliferation in liquid medium was more efficient compared with proliferation in semi-solid medium, providing a multiplication rate of 3.12 in an 8-day subculture period. As a necessary exogenous plant growth regulator (PGR) for somatic embryo maturation in conifers, abscisic acid (ABA) was optimized at 16 mg L−1 in this system. Next, an orthogonal test on osmotic pressure factors showed 50 g L−1 sucrose, 7 g L−1 phytagel and 75 g L−1 polyethylene glycol (PEG) was the optimal combination for somatic embryo maturation in L. principis-rupprechtii Mayr. Moreover, the dispersion culture method provided a more efficient somatic embryo maturation, up to 545 per gram of fresh weight (FW). Finally, 2 g L−1 of active charcoal (AC) was found to increase the somatic embryo germination rate to 63.46%. The improved protocol of SE will serve as a foundation for establishing mass propagation and genetic transformation of L. principis-rupprechtii Mayr.


Plant Science ◽  
2016 ◽  
Vol 253 ◽  
pp. 98-106 ◽  
Author(s):  
Hugo Pacheco de Freitas Fraga ◽  
Leila do Nascimento Vieira ◽  
Catarina Corrêa Puttkammer ◽  
Henrique Pessoa dos Santos ◽  
Julio de Andrade Garighan ◽  
...  

1995 ◽  
Vol 43 (4) ◽  
pp. 385-390 ◽  
Author(s):  
S. Kulothungan ◽  
A. Ganapathi ◽  
A. Shajahan ◽  
K. Kathiravan

Embryogenic callus was induced from seedling leaf explants of cowpea (Vigna unguiculata (L.) Walp. cv. C152 on Murashige and Skoog (MS) medium containing 2.0 mg 1−1 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of somatic embryogenesis was noticed when this callus was transferred to MS liquid medium supplemented with 2 mg 1−1 2,4-D. Further studies on ontogeny of somatic embryos showed that the cells destined to become somatic embryos divided into spherical or filamentous proembryos. Subsequent divisions in the proembryo led to globular, heart, torpedo-shaped, and cotyledonary-stage somatic embryos. Tiny plantlets were obtained by transferring the cotyledonary-stage somatic embryos to MS liquid medium containing 0.5 mg 1−1 2,4-D.


2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).


2018 ◽  
Vol 86 (2) ◽  
Author(s):  
Rizka Tamania SAPTARI ◽  
. SUMARYONO

 Date palm (Phoenix dactylifera L.) is the most important crop in the dry areas of the Middle East and North Africa. This palm has been introduced to many countries but has not been grown commercially in Indonesia. Date palm propaga-tion by seeds is easy but its progenies are varied and a half of them are male trees that will not produce fruits. Meanwhile, the propagation by offshoots is impractical and technically difficult. Tissue culture makes it possible to massproduce of genetically identicalsuperior date palms. This research aimed to develop somatic embryogenesis (SE) of date palm using shoot tipand young leaves of date palm seedling as explants. Steps on somatic embryogenesis are explant sterilization, callus initiation and proliferation, somatic embryos induction and maturation, and plantlets matura-tion and rooting. Calli emerged from shoot tip explants after  9 weeks of culture in a modified MS medium supplemented with 10 mg/L 2,4-D, 1 mg/L or  3 mg/L 2-iP, and 1.5 g/L active charcoal. The callus was able to bear somatic embryo in the modified MS medium without hormones. Somatic embryos then developed into plantlets, and roots of plantlets were effectively initiated in the medium supplemented with 0.5 mg/L NAA and 1 mg/L IBA.[Keywords:sterilization,  callogenesis, somatic embryo induction, plantlet rooting, clonal propagation]. Abstrak  Tanaman kurma (Phoenix dactyliferaL.) merupakan tanaman terpenting di wilayah kering Timur Tengah dan Afrika Utara. Palma ini telah menyebar ke banyak negara, namun belum ditanam secara komersial di Indonesia. Perbanyakan kurma dengan biji sangat mudah tetapi turunannya sangat beragam dan setengahnya merupakan tanaman jantan yang tidak berbuah. Perbanyakan dengan anakan (offshoots) secara komersial tidak praktis dan relatif sulit. Kultur jaringan memungkinkan untuk dihasilkan secara massal bibit tanaman kurma varietas unggul yang secara genetik seragam. Penelitian ini bertujuan untuk mengembangkan embriogenesis somatik menggunakan eksplan pucuk tunasdan daun muda dari bibit tanaman kurma. Pengembangan embriogenesis somatik terdiri dari tahap sterilisasi eksplan, inisiasi dan proliferasi kalus, induksi dan maturasi embrio somatik, serta pembesaran dan pembentukan akar planlet. Kalus terbentuk dari eksplan pucuk tunassetelah 9 minggu dikultur pada medium MS modifikasi yang ditambahkan 2,4-D 10 mg/L,  2-iP 1 mg/L atau 3 mg/L, dan arang aktif 1,5 g/L.Kalus berhasil diinduksi menghasilkanembrio somatik pada medium MS modifikasi tanpa penggunaan hormon. Embrio somatik kemudian berkembang hingga menjadi planlet, dan akar planlet secara efektif terinisiasipada medium yang ditambahkan NAA 0,5 mg/L dan IBA1 mg/L.  [Kata kunci :sterilisasi,  kalogenesis, induksi embrio somatik, pengakaran planlet, propagasi klonal].


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