scholarly journals Plant Regeneration via Somatic Embryogenesis from Leaf and Floral Explants of ‘Chancellor’ Wine Grape

1970 ◽  
Vol 20 (2) ◽  
pp. 157-170 ◽  
Author(s):  
Richard M.S. Mulwa ◽  
Margaret M.A. Norton ◽  
Robert M. Skirvin

Abundant embryogenic callus was obtained from leaf and floral explants of "Chancellor" grape by continuous culture for 12 weeks on Nitsch and Nitsch basal medium supplemented with 9 μM 2, 4-D + 17 μM IASP + either 1 μM BA or 1 μM TDZ (ECIM) in darkness. They were successfully maintained by a five to six week subculture interval on NN medium containing 2 μM 2, 4-D + 0.2 μM TDZ + 4 μM IASP (LTMM). Near synchronous embryo developed from embryogenic callus on medium containing 10 μM IASP + 8 μM NOA + 1 μM TDZ + 1 μM ABA + 2.5 g/l AC (EDMM).  Individually separated somatic embryos were germinated on both NN and half strength of MS containing 0.5 μM BA + 0.025 μM NAA, respectively; normal plantlet conversion from embryos was low (35%).  Whole fruiting plants were obtained. Aberrant embryo development was characterized by failure to form functional shoot meristems following the initial cotyledon expansion during germination. These observations indicate that the embryo conversion stage of the regeneration is difficult and remains a limiting factor requiring more empirical experimentation for improvement in grape tissue culture.   Key words: Chancellor grape, Regeneration, Somatic embryogenesis   D.O.I. 10.3329/ptcb.v20i2.6895   Plant Tissue Cult. & Biotech. 20(2): 157-170, 2010 (December)

2006 ◽  
Vol 1 (3) ◽  
pp. 1934578X0600100
Author(s):  
Bishnu P. Chapagain ◽  
Vinod Saharan ◽  
Dan Pelah ◽  
Ram C. Yadav ◽  
Zeev Wiesman

This study describes the effects of plant growth regulators, explants, and somatic embryogenesis on in vitro production of the steroidal sapogenin, diosgenin, in callus cultures of the Balanites aegyptiaca (L.) Del.(desert date). Root, shoot, hypocotyl, and epicotyl callus culture of B. aegyptiaca, were raised on MS basal media supplemented with various combinations of either 2,4-D and NAA alone, or with BAP. The diosgenin content (on a dry weight basis) was found to be highest when calli were cultured in MS basal medium supplemented with 1.0 mg l−1 2,4-D alone and/or in combination with 0.5 mg l−1 BAP. However, the callus growth was highest in media supplemented with 2.5 or 3.0 mg l−1 2,4-D. MS basal media supplemented with 2,4-D 2.5 mg l−1 alone and in combination with 0.5 mg l−1 BAP induced pre-embryogenic callus formation on root cultures. When these pre-embryogenic callus cultures were used to establish cell suspension cultures, two growth densities were obtained in embryogenic suspension cultures, inducing clusters of somatic embryos at various stages of development. The maximum number of somatic embryos were obtained at the fifth week on the medium supplemented with 1.0 mg l−1 2,4-D. However, the diosgenin content in these somatic cells was found to be lower compared to the explant calluses. This study revealed that production of diosgenin in callus cultures of B. aegyptiaca is possible, but the amount is significantly affected by the growth regulators, type of explants, and somatic embryogenesis.


1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)


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>


Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 168
Author(s):  
Muhammad Ajmal Bashir ◽  
Cristian Silvestri ◽  
Amelia Salimonti ◽  
Eddo Rugini ◽  
Valerio Cristofori ◽  
...  

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.


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.


2020 ◽  
Vol 20 (4) ◽  
pp. 179 ◽  
Author(s):  
NUR AJIJAH

<p>ABSTRAK</p><p><br />Embriogenesis somatik kakao (Theobroma cacao L.) telah banyak<br />dilaporkan  dengan  penggunaan  zat  pengatur  tumbuh  (ZPT)  yang<br />bervariasi. Penggunaan thidiazuron untuk menginduksi embriogenesis<br />somatik kakao telah dilaporkan melalui dua tahap induksi kalus. Penelitian<br />ini bertujuan untuk mengevaluasi efektivitas thidiazuron menginduksi<br />embriogenesis somatik kakao melalui satu tahap induksi kalus. Penelitian<br />dilaksanakan di Laboratorium Kultur Jaringan Unit Pengembangan Benih<br />Unggul, Badan Litbang Pertanian, Bogor. Empat taraf thidiazuron (0; 2,5;<br />5,0; dan 10 µg/l) dikombinasikan dengan 2,4-D 2 mg/l<br />digunakan untuk<br />menginduksi kalus dan embrio somatik 3 klon kakao (TSH858, Sca6, dan<br />ICS13) menggunakan eksplan mahkota bunga dan staminoid. Media dasar<br />DKW tanpa ZPT digunakan sebagai kontrol. Penelitian disusun dalam<br />rancangan lingkungan acak lengkap dengan lima ulangan. Setiap unit<br />percobaan terdiri dari sepuluh eksplan. Peubah yang diamati meliputi<br />persentase pembentukan kalus umur 2 dan 4 minggu, penampakan visual<br />kalus, persentase eksplan membentuk embrio somatik, dan jumlah embrio<br />somatik per eksplan umur 10 dan 14 minggu. Kalus terbentuk pada media<br />dengan penambahan hanya 2,4-D atau 2,4-D + thidiazuron, namun embrio<br />somatik hanya terbentuk pada media dengan penambahan 2,4-D +<br />thidiazuron. Pembentukan kalus dan embrio somatik sangat dipengaruhi<br />oleh tipe eksplan dan genotipe. Klon Sca6 lebih responsif dibandingkan<br />TSH858 dan ICS13 dan eksplan staminoid lebih responsif dibandingkan<br />mahkota bunga. Hasil studi ini menunjukkan terdapat pengaruh interaksi<br />yang kuat antara ZPT, genotipe, dan tipe eksplan terhadap pembentukan<br />kalus dan embrio somatik kakao serta tidak terdapat perbedaan hasil yang<br />nyata antara pembentukan embrio somatik melalui satu dan dua tahap<br />induksi kalus.<br />Kata kunci: Theobroma cacao L., genotipe, eksplan, zat pengatur tumbuh</p><p>ABSTRACT</p><p><br />Somatic embryogenesis of cacao (Theobroma cacao L.) has been<br />widely reported with varied of plant growth regulators (PGR) used. The<br />use of thidiazuron in inducing somatic embryogenesis of cacao has been<br />reported through a two-step callus induction. The study aimed to evaluate<br />the effectiveness of thidiazuron in inducing somatic embryogenesis of<br />cacao through a one-step of callus induction. The study was conducted at<br />the tissue culture laboratory of Agricultural Seed Development Unit,<br />Indonesian Agency for Agricultural Research and Development, Bogor.<br />Four levels of thidiazuron (0; 2.5; 5.0; and 10 µg/l) in combination with 2<br />mg/l  2,4-D  were  used  for  inducing  callogenesis  and  somatic<br />embryogenesis of three cacao clones (TSH858, Sca6, and ICS13) using<br />petals and staminoids explants. DKW basal medium without PGR was<br />used as a control. The result showed that callus were formed on medium<br />containing only 2,4-D or 2,4-D + thidiazuron, while embryos were only<br />formed on medium containing 2,4-D + thidiazuron. The formation of<br />callus and somatic embryos were highly affected by explant types and<br />genotypes. Sca6 clone was more responsive than TSH858 and ICS13 and<br />staminoids were more responsive than petals. The results of this study<br />revealed that there was a strong interaction between the PGRs, genotypes,<br />and explant types on the formation of cacao callus and somatic embryos.<br />Results of this study also showed no significant difference between the<br />formation of somatic embryos through one and two steps of callus<br />induction.<br />Keywords: Theobroma cacao L., genotypes, explants, plant growth<br />regulators</p>


2021 ◽  
Vol 306 ◽  
pp. 01056
Author(s):  
Sulistyani Pancaningtyas

Somatic embryogenesis is one of the newest technology that applied for the mass production of cocoa. This research aims to evaluate the regeneration rate of somatic embryos through somatic embryogenesis propagation techniques on java fine flavor cocoa. Cultivars in this study are ICCRI 01, ICCRI 02, DR 1, DR 2, DRC 16, DR 38, PNT 16, and PNT 30. Observations include parameters to determine the percentage of primary callus and embryogenic callus formation and the number of somatic embryos produced. Based on data, the ability of callus to produce primary embryos is highly dependent on plant cultivars and explant sources. Five cultivars showed a higher regeneration rate using explants from the petal part, while the rest showed a higher regeneration rate using explants from the staminode section. Embryogenic callus from each cacao cultivar has the same basic structure: a nodular friable structure consisting of many embryonic cells. Some fine flavor cacao cultivars that were able to produce callus and primary somatic embryos could not produce secondary somatic embryos and plantlets. However, two cultivars, which had low potential in producing primary embryos, had the high ability to produce secondary somatic embryos and develop into plantlets.


PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5952 ◽  
Author(s):  
Guadalupe Hernández-Piedra ◽  
Violeta Ruiz-Carrera ◽  
Alberto J. Sánchez ◽  
Arlette Hernández-Franyutti ◽  
Alfonso Azpeitia-Morales

Background Sustainable methods of propagation of Typha domingensis through somatic embryogenesis can help mitigate its current condition of ecological marginalization and overexploitation. This study examined whether differentiation up to coleoptilar embryos could be obtained in an embryogenic line proliferated with light and high auxin concentration. Methods Murashige and Skoog medium at half ionic strength and containing 3% sucrose and 0.1% ascorbic acid was used for the three embryogenic phases. Induction started with aseptic 9-day-old germinated seeds cultured in 0.5 mg L−1 2,4-dichlorophenoxyacetic (2,4-D). Proliferation of the embryogenic callus was evaluated at 2,4-D concentrations ranging from 0 to 2 mg L−1 in cultures maintained in the dark. The dominant embryogenic products obtained in each treatment were used as embryogenic lines in the third phase. Thus, maturation of the somatic embryos (SEs) was analyzed using four embryogenic lines and under light vs. dark conditions. Embryogenic differentiation was also monitored histologically. Results Proliferation of the nine morphogenetic products was greater in the presence of 2,4-D, regardless of the concentration, than in the absence of auxin. Among the products, a yellow callus was invariably associated with the presence of an oblong SE and suspended cells in the 2,4-D treatments, and a brown callus with scutellar somatic embryos (scSEs) in the treatment without 2,4-D. During the maturation phase, especially the embryogenic line but also the light condition resulted in significant differences, with the highest averages of the nine morphogenetic products obtained under light conditions and the maximum concentration of auxin (YC3 embryogenic line). Only this line achieved scSE growth, under both light and dark conditions. Structurally complete coleoptilar somatic embryos (colSEs) could be anatomically confirmed only during the maturation phase. Discussion In the embryogenic line cultured with the highest auxin concentration, light exposure favored the transdifferentiation from embryogenic callus to scSE or colSE, although growth was asynchronous with respect to the three embryogenic phases. The differentiation and cellular organization of the embryos were compatible with all stages of embryogenic development in other monocotyledons. The growth of colSEs under light conditions in the YC3 embryogenic line and the structurally complete anatomic description of colSEs demonstrated that differentiation up to coleoptilar embryos could be obtained. The diversity of embryogenic products obtained in the YC3 embryogenic line opens up the opportunity to synchronize histological descriptions with the molecules associated with the somatic embryogenesis of Typha spp.


HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 629g-630 ◽  
Author(s):  
S. Jayasankar ◽  
U.L Yadava

Petiole discs from young leaves of female papaya (L-45) plants were cultured in MS or B5-based media containing 0, 2.25, 4.5, 11.25, and 22.5 μm 2,4-D. Compact embryogenic callus emerged from vascular tissue of petiole discs in about 3 weeks. In MS medium, 66% and 51% explants formed embryogenic callus with 11.25 and 22.5 μm 2,4-D, respectively. On the other hand, 79% explants formed embryogenic callus in B5-based medium with 4.50 μm 2,4-D. However, explants became necrotic in B5-based medium with 22.5 μm 2,4-D. Subculturing callus in auxin-free medium resulted in the development of roots or somatic embryos. Microscopic observations revealed that the roots were produced only by the callus that had retained its continuity with the vascular tissue. This investigation revealed that petioles from field grown papaya plants are potential explants for somatic embryogenesis and 2-week exposure to 2,4-D is adequate for inducing morphogenesis. Additionally, an interaction between 2,4-D and the components in the MS and B5-based media was observed.


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