PLANT REGENERATION FROM CALLUS CULTURES DERIVED FROM MATURE ZYGOTIC EMBRYOS OF SOPHORA ALOPECUROIDES LINN. IN MONGOLIA

Author(s):  
Tsolmon M ◽  
Ganbat B ◽  
Oyunbileg Yu

The aim of this study is to determine the effect of hormones and selection of the most effective medium using callus cultures derived from mature zygotic embryos of Sophora alopecuroides Linn. for plant regeneration. After 8 weeks of culture, the highest callus induction medium (93.3%) was obtained on MS medium supplemented with 0.2 mglL Zeatin and 2.0 mg/L α-naphthaleneacetic acid (NAA). The best callus proliferation was observed on the same medium. Shoots regenerated at the highest frequency of 50.0% with 5.8 shoots when calli were cultured on MS medium with 2.0 mg/L BA. Therefore, this protocol provides a basis for future studies on genetic improvement and could be applied to large-scale multiplication systems for commercial nurseries of S.alopecuroides L.

HortScience ◽  
1991 ◽  
Vol 26 (10) ◽  
pp. 1322-1324 ◽  
Author(s):  
A. Levi ◽  
K.C. Sink

The histology and morphology of developing asparagus Asparagus officinalis L.) somatic embryos arising in callus cultures were examined and contrasted with that documented for zygotic embryos. Histological sections of lateral bud-derived callus cultured for 2 weeks on embryo induction medium consisting of Murashige and Skoog salts and vitamins (MS) with 1.5 mg NAA/liter and 0.1 mg kinetin/liter indicated the formation of distinct groups of embryogenic cells. At 4 weeks, the callus was comprised of embryos in the early and late globular stages and a few bipolar embryos. Within 2 weeks on embryo development medium consisting of MS with 0.05 mg NAA/liter and 0.1 mg kinetin/liter, the globular embryos developed a bipolar shape having an expanded upper region that formed the cotyledon and a smaller region that formed the radicle. Within 4 to 6 weeks on this latter medium, each mature bipolar embryo was opaque and had a large cotyledon, a distinct shoot apex at the cotyledon-hypocotyl junction, and vascular connections between the radicle, shoot apex, and cotyledon. Many mature somatic embryos resembled the asparagus zygotic embryos in having a crescent shape, whereas others had a short but wide cotyledon. Both somatic embryo types converted to plantlets at equal rates. Chemical names used: N- (2-furanylmethyl)-1 H -purin-6-amine (kinetin); 1-naphthaleneacetic acid (NAA).


Hoehnea ◽  
2017 ◽  
Vol 44 (1) ◽  
pp. 90-95 ◽  
Author(s):  
Andressa Reis ◽  
Alítcia Moraes Kleinowski ◽  
Fátima Rosane Schuquel Klein ◽  
Renata Trevizan Telles de Souza ◽  
Luciano do Amarante ◽  
...  

ABSTRACT The objective of this study was to establish a protocol for callus induction and betacyanin production in plants of Alternanthera brasiliana. Explants of A. brasiliana and five combinations of cytokinin and auxin were used for callus induction. Calli were transferred to a Betacyanin Induction Medium (MIB), composed of MS, with 0.5 mg L-1 of thidiazuron (TDZ) and 1 mg L-1 of naphthaleneacetic acid (NAA), and kept in the light for 45 days. The aspect and intensity of pigments were assessed and total betacyanins were quantified in high performance liquid chromatography (HPLC). The combination of internodal segments and a medium containing equilibrium concentrations of auxins and cytokinins was the most efficient metod to induce calli and increased production of betacyanins. The presence of amaranthine in calli of A. brasiliana justifies its medical use and the consequent need for future studies for the large-scale production of this molecule.


Author(s):  
Masoumeh Nomani ◽  
Masoud Tohidfar

Abstract Background Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi. Results According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants. Conclusions In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25–35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.


2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis


2012 ◽  
Vol 7 (4) ◽  
pp. 680-689 ◽  
Author(s):  
Laura Bedini ◽  
Mariella Lucchesini ◽  
Francesco Bertozzi ◽  
Alberto Graifenberg

AbstractThe aim of the study was to examine the possibility of propagating in vitro four of the most common cultivars in Tuscany (central Italy): Terom, Violetto di Toscana, Chiusure and Empolese. The first three belong to the “Violetti” group, while cv Empolese belongs to the “Romaneschi” group. Explants were cultured on an induction medium (IM), which is a modified MS medium consisting of nitrate concentrations reduced by one quarter, 0.8 mg L−1 6-benzylaminopurine (BA) and 0.2 mg L−1 3-indole butyric acid (IBA). Explants were then transferred to a proliferation medium (PM) consisting of the same basal medium together with 0.03 mg L−1 BA and 0.05 mg L−1 gibberellic acid (GA3). A rooting double-phase was then established. The pre-rooting medium (PRM), consisting of a basal MS medium with half strength nitrate concentrations, 0.5 mg L−1 indole-3-acetic acid (IAA) and 1 mg L−1 paclobutrazol (PBZ) was used for two weeks. Over the next four weeks, a rooting medium (MR) was used, consisting of a basal MS medium with 2 mg L−1 β-cyclodextrin and 2 mg L−1 α-naphthaleneacetic acid sodium salt (NAA). The cv Empolese provided the highest number of proliferated explants and rooted plantlets using the method described.


HortScience ◽  
2017 ◽  
Vol 52 (9) ◽  
pp. 1278-1282 ◽  
Author(s):  
Boling Liu ◽  
Hongzhou Fang ◽  
Chaorong Meng ◽  
Ming Chen ◽  
Qingdong Chai ◽  
...  

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.


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


2019 ◽  
Vol 20 (9) ◽  
pp. 2141 ◽  
Author(s):  
Han Yong Park ◽  
Doo Hwan Kim ◽  
Ramesh Kumar Saini ◽  
Judy Gopal ◽  
Young-Soo Keum ◽  
...  

The goal of this study was to establish an efficient protocol for the large-scale propagation of Mertensia maritima (L.) Gray, and evaluate the carotenoid, fatty acid, and tocopherol contents in the leaves of in vitro regenerated shoots. Surface-disinfected node and shoot tip explants were placed on semisolid Murashige and Skoog (MS) medium with 0–16 µM N6-benzyladenine (BA), kinetin, (KN), and thidiazuron (TDZ) alone, or in combination with, 1 or 2 µM α-naphthaleneacetic acid (NAA). Of the three different cytokinins employed, TDZ elicited the best results for axillary shoot proliferation. A maximum frequency of shoot initiation above 84%, with a mean of 8.9 and 4.8 shoots per node and shoot tip, respectively, was achieved on the culture medium supplemented with 4 µM TDZ. A combination of TDZ + NAA significantly increased the percentage of multiple shoot formation and number of shoots per explant. The best shoot induction response occurred on MS medium with 4 µM TDZ and 1 µM NAA. On this medium, the node (93.8%) and shoot tip (95.9%) explants produced an average of 17.7 and 8.6 shoots, respectively. The highest root induction frequency (97.4%) and number of roots per shoot (25.4), as well as the greatest root length (4.2 cm), were obtained on half-strength MS medium supplemented with 4 µM indole-3-butyric acid (IBA). The presence of six carotenoids and α-tocopherol in the leaf tissues of M. maritima was confirmed by HPLC. Gas chromatography-mass spectrometry analysis confirmed the presence of 10 fatty acids, including γ-linolenic acid and stearidonic acid in the leaf tissues of M. maritima. All-E-lutein (18.49 μg g−1 fresh weight, FW), α-tocopherol (3.82 μg g−1 FW) and α-linolenic acid (30.37%) were found to be the significant compounds in M. maritima. For the first time, a successful protocol has been established for the mass propagation of M. maritima with promising prospects for harnessing its bioactive reserves.


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.


HortScience ◽  
2014 ◽  
Vol 49 (12) ◽  
pp. 1558-1562 ◽  
Author(s):  
Yuyu Wang ◽  
Faju Chen ◽  
Yubing Wang ◽  
Xiaoling Li ◽  
Hongwei Liang

High-frequency somatic embryogenesis and plant regeneration were achieved from immature cotyledonary-stage embryos in the endangered plant, Tapiscia sinensis Oliv. Plant growth regulators with different concentrations and combinations on embryogenesis capacity were studied. The optimal explants for in vitro somatic embryogenesis were immature embryos in T. sinensis. A high callus induction rate of 100% was achieved on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg·Ll−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5% (w/v) activated charcoal. Alternatively, a high induction rate (96.16%) of somatic embryogenesis was obtained on MS basal medium supplemented with the combination of 0.05 mg·L−1 α-naphthaleneacetic acid (NAA) and 0.2 mg·L−1 6-benzylaminopurine (6-BA), and somatic embryos proliferated fastest on the mentioned medium supplemented with 0.5% (w/v) activated charcoal and 3% (w/v) sucrose, inoculation of explants proliferating 21 times in the 23-day subculture. Of the 100 plantlets transferred to field after the acclimation, 95 (95%) survived. Based on the histocytological observations, the development of somatic embryos was similar to that of zygotic embryos. There were two accumulation peaks of starch grains in the embryogenic calli and in the globular-stage embryos, both closely related to the energy supply, and the embryoids were of multicelluar origin.


Sign in / Sign up

Export Citation Format

Share Document