scholarly journals The Origin of the Cyathea delgadii Sternb. Somatic Embryos Is Determined by the Developmental State of Donor Tissue and Mutual Balance of Selected Metabolites

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1388
Author(s):  
Anna Mikuła ◽  
Wojciech Tomaszewicz ◽  
Michał Dziurka ◽  
Andrzej Kaźmierczak ◽  
Małgorzata Grzyb ◽  
...  

Somatic embryogenesis is the formation of a plant embryo from a cell other than the product of gametic fusion. The need to recognize the determinants of somatic cell fate has prompted investigations on how endogenous factors of donor tissues can determine the pattern of somatic embryo origin. The undertaking of this study was enabled by the newly developed experimental system of somatic embryogenesis of the tree fern Cyathea delgadii Sternb., in which the embryos are produced in hormone-free medium. The contents of 89 endogenous compounds (such as sugars, auxins, cytokinins, gibberellins, stress-related hormones, phenolic acids, polyamines, and amino acids) and cytomorphological features were compared between two types of explants giving rise to somatic embryos of unicellular or multicellular origin. We found that a large content of maltose, 1-kestose, abscisic acid, biologically active gibberellins, and phenolic acids was characteristic for single-cell somatic embryo formation pattern. In contrast, high levels of starch, callose, kinetin riboside, arginine, and ethylene promoted their multicellular origin. Networks for visualization of the relations between studied compounds were constructed based on the data obtained from analyses of a Pearson correlation coefficient heatmap. Our findings present for the first time detailed features of donor tissue that can play an important role in the somatic-to-embryogenic transition and the somatic embryo origin.

Author(s):  
Muniappan V ◽  
Manivel P ◽  
Prabakaran V ◽  
Palanivel S ◽  
Parvathi S

Somatic embryogenesis was carried out epicotyl portion of the mature embryo/apical portion. The somatic embryo induction medium containing 2,4-D or NAA (10.0 to 50.0 mg/l). Of the two concentrations tested 2,4-D (30.0mg/l) recorded the highest percentage of response followed by NAA (30.0mg/l). But the highest number of somatic embryo were recorded in 30.0mg/l of 2,4-D followed by NAA. The apical portion of the mature embryo formed direct embryos without any intervention of callus. The maximum percentage of embryogenic cultures were noticed in 30.0mg/l of 2,4-D followed by NAA at 30.0mg/l. for the differentiation of somatic embryos, the embryogenic masses were transferred to medium without any growth regulator. The maximum number of somatic embryos per culture was recorded in 30 mg/l of 2,4-D followed by 30.0 mg/l of NAA. Keywords: Arachis hypogaea L.,Somatic Embryogenesis, 2,4-D and NAA


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


1997 ◽  
Vol 75 (3) ◽  
pp. 509-512 ◽  
Author(s):  
P. V. Bozhkov ◽  
I. S. Ahn ◽  
Y. G. Park

Individual mature stored seeds of Pinus koraiensis sometimes contain several viable zygotic embryos originated through the processes of simple and cleavage polyembryony. To induce the embryonic process, isolated zygotic embryos were cultured on five different media all supplemented with 10 μM 2,4-dichlorophenoxyacetic acid and 5 μM 6-benzyladenine. Two alternative pathways of somatic embryo origin were revealed. The first pathway was associated with the production of a friable, translucent callus in the hypocotyls–cotyledon region of the dominant zygotic embryo. The second pathway was related to the proliferation of a translucent, moist, and mucilaginous tissue (termed embryonal–suspensor mass) in the suspensor region of the dominant zygotic embryo. Both types of tissues contained early somatic embryos. Regression analysis has shown a strong negative correlation between the frequencies of formation of embryogenic callus and embryonal–suspensor mass both at 3 and 8 weeks of culture (r = − 0.85; p = 0.07 and r = −0.71; p = 0.17, respectively). Key words: Pinus koraiensis; polyembryonal seeds; somatic embryogenesis; embryogénie callus; embryonal–suspensor mass.


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].


HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 251f-251 ◽  
Author(s):  
Christopher S. Cramer ◽  
Mark P. Bridgen

Disinfected midrib sections of Mussaenda `Queen Sirikit' ≈3 to 4 mm in size were cultured on a basal medium of Murashige and Skoog salts and vitamins, 87.7 mm sucrose, and 5 g Sigma agar/liter supplemented with several concentrations of indole-3-acetic acid (IAA) (0, 5.0, 10.0, 20.0 μm) and 6-benzylaminopurine (BAP) (0, 0.5, 1.0, 2.5, 5.0, 10.0, 25.0, 50.0 μm). Cultures were subculture onto the same treatment after 5 weeks and observed weekly for 15 weeks for the presence of somatic embryos. As somatic embryos were produced, they were subculture onto basal medium supplemented with 0.5, 1.0, 2.5, or 25.0 μm BAP. Callus was first observed at 2 weeks in cultures grown on basal medium supplemented with 5.0–20.0 μm IAA and 0–50.0 μm BAP. Somatic embryos were observed at 8 weeks on basal medium supplemented with 5.0–10.0 μm IAA and 2.5–5.0 μm BAP. Callus cultured on 0–10 μm IAA and 5.0–10.0 μm BAP produced the greatest number of somatic embryos by 15 weeks. Somatic embryos subculture to basal medium supplemented with 25.0 μm BAP proliferated shoots, while eliminating BAP from the medium resulted in root and callus production. Shoots and entire plants were removed from in vitro conditions and successful] y acclimated to greenhouse conditions. Somatic embryo-derived plants flowered sporadically 25 to 35 weeks after removal from in vitro conditions. Variations in sepal number and leaf number per node were observed at 1% to 5%.


2019 ◽  
Vol 49 (12) ◽  
pp. 1604-1612
Author(s):  
Tingyu Sun ◽  
Yanli Wang ◽  
Lihua Zhu ◽  
Xiaoqin Wu ◽  
Jianren Ye

Pine wilt disease (PWD) is a severe threat to pine forests in East Asia. Screening and breeding of resistant varieties is a very effective way to prevent and control PWD; however, no reliable somatic embryogenesis system has yet been developed for the elite nematode-resistant Pinus thunbergii Parl. line. In this study, we studied the plant regeneration via somatic embryogenesis of nematode-resistant P. thunbergii. Initiation of embryogenic tissue was significantly affected by seed family (p = 0.017), immature zygotic embryo stage (p = 0.032), and initiation medium (p = 0.004). Seed family 37 was the most favorable female parent for initiation of P. thunbergii. Furthermore, the initiation rate increased from the pre-embryonic stage to the cleavage polyembryonic stage. The optimal medium was I2, containing 2,4-dichlorophenoxyacetic acid (9 μmol·L−1) and 6-benzyladenine (4.4 μmol·L−1). A statistically significant interaction between cell line and subculture time (24 months) was observed in the influence on proliferation rate, somatic embryo production, and percentage germination (p < 0.001). In this study, the highest somatic embryo production was achieved using cell line 37-1 (1983 somatic embryos per gram fresh mass), with approximately 83.5% of somatic embryos germinating after transferring to germination medium, of which 77.6% converted into plantlets.


2016 ◽  
Vol 2 (3) ◽  
pp. 6-13 ◽  
Author(s):  
Meynarti Sari Dewi Ibrahim ◽  
Raden Roro Sri Hartati ◽  
Rubiyo Rubiyo ◽  
Agus Purwito ◽  
Sudarsono Sudarsono

The primary and secondary somatic embryogenesis can be used to propagate Coffea arabica L clonally.  However, the success of this propagation was depended on plant growth regulator and varieties. This study aimed to examine the possibility of 2,4-D and thidiazuron application to form primary and secondary somatic embryo to support Arabica coffee clonal propagation. The study consisted of two activities (1) 2,4-D and thidiazuron Application to Induce Primary Somatic Embryogenesis of Arabica Coffee and (2) The Application of thidiazuron in Solid and Semi-Solid Media to Induce Secondary Somatic Embryos.  The results indicated significant effect of varieties and plant growth regulator on fresh weight, number of torpedo and germinated embryo.  However, it showed no significant effect on callus formation percentage. The best medium to induce primary somatic embryogenesis depending on variety, on the treatment of 4.52 μM 2,4 -D +18.16 μM thidiazuron was the best for AS2K and Sigarar Utang varieties, S 795 at 4.52 μM 2,4-D + 9.08 μM thidiazuron, whereas Kartika at 4.52 μM 2.4-D + 13.62 μM thidiazuron.  The morphology of coffee somatic embryo was normal.  Primary somatic embryo was developed indirectly, whereas the secondary somatic embryo was directly.  The application of 9.08 μM thidiazuron  increased the percentage and number of secondary somatic embryos, hence enhancing number of Arabica coffee planlet. Keywords : Coffea arabica L, 2,4-D, thidiazuron, semi-solid media, Indirect somatic embryogenesis


2021 ◽  
Author(s):  
Claudia Garcia ◽  
Alex-Alan Furtado de Almeida ◽  
Marcio Costa ◽  
Dahyana Britto ◽  
Fabio Correa ◽  
...  

Abstract Propagation by somatic embryogenesis in Theobroma cacao has some issues to be solved, as many morphologically abnormal somatic embryos that do not germinate into plants are frequently observed, thus hampering plant production on a commercial scale. For the first time the methylome landscape of T. cacao somatic embryogenesis was examined, using whole-genome bisulfite sequencing technique, with the aim to understand the epigenetic basis of somatic embryo abnormalities. We identified 873 differentially methylated genes (DMGs) in the CpG context between zygotic embryos, normal and abnormal somatic embryos, with important roles in development, programmed cell death, oxidative stress, and hypoxia induction, which can help to explain the morphological abnormalities of somatic embryos. We also identified the role of ethylene and its precursor 1-aminocyclopropane-1-carboxylate in several biological processes, such as hypoxia induction, cell differentiation and cell polarity, that could be associated to the development of abnormal somatic embryos. The biological processes and the hypothesis of ethylene and its precursor involvement in the somatic embryo abnormalities in cacao are discussed.


HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 568B-568a
Author(s):  
Lianghong Chen ◽  
Ajmer S. Bhagsari ◽  
Soon O. Park ◽  
Sarwan Dhir

This study was carried out to optimize conditions for plant regeneration of sweetpotato [Ipomoea batatas (L.) Lam] using shoot tips, petioles, and leaves of Selection 75-96-1 as explants in Murashige and Skoog (MS) with several growth regulators at different levels. Callus initiation and callus proliferation media were 9.0 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and 9.0 μm 2,4-D + 1.1 μm N6-benzyladenine (6-BA) in protocol I; 8.1 μm α-naphthaleneacetic acid (NAA) + 1.2 μm kinetin (KIN) and 5.4 μm NAA + 4.6 μm KIN in protocol II; 0.9 μm 2,4-D, and 0.9 μm 2,4-D + 1.2 μm N-isopenylamino purine (2iP) in protocol III; NAA (8.1 μm) + KIN (1.2 μm) and 2,4-D (0.9 μm) + 2ip (1.2 μm) in protocol IV, respectively. In protocol I and II, shoot tip, petiole, and leaf were used, but only petiole and leaf in protocol III and IV. In the protocol I and II, somatic embryos were obtained only from shoot tip explants; in protocol III and IV, only from petioles. The frequencies of somatic embryo development were 33.3% in protocol I, 42.1% in protocol II, 21.2% in protocol III, and 10.3% in protocol IV, respectively. The leaf explants failed to produce somatic embryos in all the experiments. In protocol I, somatic embryogenesis occurred through the well-known sequence of globular-, heart-shaped-, torpedo-, and cotyledon-type embryos. However, in protocol II, the structures resembling plumule and radicle were observed before the emergence of torpedo/cotyledon type embryo clusters. The somatic embryogenesis in protocol III and IV was similar to that in protocol I. Growth regulators influenced somatic embryo development. Further, this study showed that explant resource and growth regulators affected the frequency of plant regeneration in sweetpotato.


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