scholarly journals Micropropagation of some Onobrychis species through in vitro shoot regeneration

2020 ◽  
Vol 19 (5) ◽  
pp. 45-52
Author(s):  
Satı Uzun ◽  
Duran YÜKSELGÜNGÖR

Onobrychis species have an extensive spread in Turkey and adapted to various environmental conditions. In this study, it was aimed to propagate with tissue culture methods of endemic Onobrychis fallax Freyn & Sint. ex Freyn var. longifolia Aktoklu, Onobrychis stenostachya subsp. sosnowskyi, Onobrychis elata Boiss. & Balansa and worldwide commonly cultivated Onobrychis viciifolia Scop. species. For this purpose, cotyledon node explants were cultured in Murashige and Skoog (MS) media containing various concentration of 6-benzylaminopurine (BAP), thidiazuron (TDZ) or meta-Topolin (mT) alone or in combination with 0.5 mg L–1 NAA. Significant variations in number of shoots per explant was observed depending on the species and growth regulators. The greatest number of shoots per explant was obtained from 1.2 mg L–1 TDZ treatments in O. stenostachya, O. fallax and O. elata and from 0.3 mg L–1 TDZ treatments in O. viciifolia, whereby 10.58, 9.50, 5.5, 6.42 shoots were recorded. Resultant shoots were rooted in half strength MS nutrient media containing 1 or 2 mg L–1 indole-3-butyric acid (IBA). The rooting ratio was 86.11% in O. viciifolia, 50.00% in O. stenostachya and 36.11% in O. fallax and O. elata.

2006 ◽  
Vol 12 (1) ◽  
Author(s):  
M. Ördögh ◽  
E. Jámbor-Benczúr ◽  
A. Tilly Mándy ◽  
L. Lelik

The Hungarian cultivar Sorbus redliana 'Burokvölgy' was proliferated on Murashige and Skoog (MS, 1962) medium with half-strength macroelements and 100 mg/1 meso-inositol, 20 g/1 sucrose, 11 g/1 agar-agar. Different combinations of kinetin (KIN), metatopolin (mT), benzyladenine (BA), benzyladenine-ribosid (BAR) and indolebutiric acid (IBA) were tested, and pH was adjusted to 5.6 every case using KOH. The cultures were incubated at 20-24 °C in 8/16 hours dark/light photoperiod for 50-52 days. The main aim of our research was to find the optimal growth regulator and its optimum concentration. Purthermore, to determine the chlorophyll contents of the in vitro propagated plants' leaves. During the proliferation, the highest number of shoots were observed in the case of using BA + IBA, and on the medium containing 0.75 mg/I BA + 0.05 mg/1 IBA 8.93 shoots were found. The addition of KIN + IBA decreased the number of shoots and increased the sizes of leaves — the widest (11.2 mm) and longest (17.8 mm) leaves were obtained on the medium containing 1.00 mg/I KIN + 0.05 mg/1 IBA. The longest shoots (36.46 mm) were found in the case of applying 0.75 mg/1 BAR + 0.05 mg/I IBA. The BA + KIN + IBA combination resulted the shortest shoots. Sometimes not only shoot regeneration but spontaneous rooting was observed during the multiplication. The highest chlorophyll content (1.569 mg/g total chlorophyll, 1.132 mg/g chlorophyll-a, 0.437 mg/g chlorophyll-b) was obtained in the presence of 1.0 mg/I KIN + 0.05 mg/1 IBA.


OENO One ◽  
2015 ◽  
Vol 49 (1) ◽  
pp. 37 ◽  
Author(s):  
Nadra Khan ◽  
Maqsood Ahmed ◽  
Ishfaq Hafiz ◽  
Nadeem Abbasi ◽  
Shaghef Ejaz ◽  
...  

<p style="text-align: justify;"><strong>Aim</strong>: To optimize the concentrations of growth regulators in the media for the proficient micropropagation of grapevine (<em>Vitis vinifera </em>L.) cv. King’s Ruby.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Apical meristems of the grape cultivar were used to establish <em>in vitro</em> shoot cultures. Nodal explants, each containing an axillary bud, taken from <em>in vitro</em> grown shoots were inoculated in shoot proliferation medium, i.e., half strength Murashige and Skoog (MS) medium supplemented with benzyl aminopurine (BAP), kinetin, glycine and gibberellic acid (GA<sub>3</sub>). A higher number of shoots (5.33) with greater shoot length (2.75 cm) was produced in the medium supplemented with 1.0 mg L<sup>-1</sup> BAP and 0.1 mg L<sup>-1</sup> GA<sub>3</sub>. Calluses were induced from leaf explants taken from <em>in vitro</em> grown shoots. Callus induction was greater (73.00%) on the medium containing 2.0 mg L<sup>-1</sup> 2,4-dichlorophenoxyacetic acid (2,4-D), 0.3 mg L<sup>-1</sup> BAP and 0.2 mg L<sup>-1</sup> α-naphthaleneacetic acid (NAA). The maximum frequency of shoot regeneration (53.33%) was achieved on the medium supplemented with 1.5 mg L<sup>-1</sup> BAP and 0.5 mg L<sup>-1</sup> NAA, and the regenerated shoots successfully formed roots on growth regulator-free half strength MS medium.</p><p style="text-align: justify;"><strong>Conclusion</strong>: Optimizing the concentration of BAP and GA<sub>3</sub> and omitting the glycine and kinetin in the culture medium increased the number and length of shoots. Similarly, for inducing the callus of the leaf explants, taken from <em>in vitro</em> grown shoots, it is recommended to adjust the medium with the higher concentration of 2,4-D and lower concentrations of BAP. Moreover, the maximum number of shoots was regenerated on a medium supplemented with relatively high levels of both BAP and NAA (1.5 and 0.5 mg L<sup>-1</sup>, respectively). Finally, we suggest the half strength MS medium that is free from growth regulators for the root formation of the regenerated shoots.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: Optimizing the concentration of growth regulators is crucial for the efficient micropropagation of a grape cultivar. Knowing the specific balance between the growth regulators is necessary to establish <em>in vitro</em> shoot cultures, callus induction and shoot regeneration and, hence, to propagate disease-free true to type grape cultivars in a short time.</p>


HortScience ◽  
2009 ◽  
Vol 44 (2) ◽  
pp. 523-525 ◽  
Author(s):  
Kazuhiko Mitsukuri ◽  
Takaya Arita ◽  
Masahumi Johkan ◽  
Satoshi Yamasaki ◽  
Kei-ichiro Mishiba ◽  
...  

Habenaria radiata is a terrestrial orchid with beautiful bird-shaped petals. The wild H. radiata population has been severely affected by environmental disruption and overexploitation. In micropropagation of H. radiata, although aseptic germination has been studied, tissue culture methods have not yet been established. Shoot apexes and leaf explants from vegetative plants and flower stalks, stolons, and floret explants from reproductive plants were chosen for this study. Explants were cultured on half-strength inorganic salts and full-strength vitamins of Murashige and Skoog (1/2 MS) medium containing 30 g·L−1 sucrose, 8 g·L−1 agar (pH 5.6) supplemented with 4.44 μM N6-benzyladenine, and 0.54 μM α-naphthaleneacetic acid. After 8 weeks of culture, the highest survival rate was obtained with floret explants excised from plants at the reproductive phase. In floret culture, the number of adventitious bud formation per explant was 5.4 per upper floret and 4.0 per lower floret. Dark preconditioning, which inhibited browning and contamination, of explants before shoot apex culture increased survival rates of explants (53%) and bud formation (83%). Consequently, a tissue culture method using florets and shoot apexes as explant material was established for H. radiata.


2009 ◽  
Vol 36 (No. 4) ◽  
pp. 140-146 ◽  
Author(s):  
J.K. Kanwar ◽  
S. Kumar

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in <I>Dianthus caryophyllus</I> L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.


2011 ◽  
Vol 3 (3) ◽  
pp. 97-100
Author(s):  
Naimeh SHARIFMOGHADAM ◽  
Abbas SAFARNEJAD ◽  
Sayed Mohammad TABATABAEI

The Almond (Amygdalus communis) is one of the most important and oldest commercial nut crops, belonging to the Rosaceae family. Almond has been used as base material in pharmaceutical, cosmetic, hygienically and food industry. Propagation by tissue culture technique is the most important one in woody plants. In the current research, in vitro optimization of tissue culture and mass production of almond was investigated. In this idea, explants of actively growing shoots were collected and sterilized, then transferred to MS medium with different concentrations and combinations of plant growth regulators. The experiment was done in completely randomized blocks design, with 7 treatment and 30 replications. After 4 weeks, calli induction, proliferation, shoot length and number of shoot per explants were measured. Results showed that the best medium for shoot initiation and proliferation was MS + 0.5 mg/l IAA (Indol-3-Acetic Acid) + 1 mg/l BA (Benzyl Adenine). Autumn was the best season for collecting explants. The shoots were transferred to root induction medium with different concentrations of plant growth regulators. The best root induction medium was MS + 0.5 mg/l IBA (Indol Butyric Acid).


2009 ◽  
Vol 18 (1) ◽  
pp. 17-24
Author(s):  
Md. Nesawar Miah ◽  
Shahina Islam ◽  
Syed Hadiuzzaman

Efforts have been made to establish a protocol for direct multiple shoot regeneration from both in vitro grown seedlings and mature plants of Citrus macroptera. Both nodal and shoot tip explants taken from in vitro grown seedlings were cultured in MS supplemented with different concentrations of BAP and Kn either singly or in combinations. Both these explants are capable to regenerate and produce in vitro multiple shoots. Maximum number of shoots were obtained from nodal explants in MS supplemented with 1.0 mg/l BAP. BAP alone was found superior to Kn. On the other hand, only nodal explants from mature plants were used and 1.0 mg/1 BAP was also found best suitable for shoot induction and multiplication. Ex vitro rooting in pot soil (mixed with biogas slurry derived from cow-dung) was most successful compared to in vitro rooting in half strength of MS supplemented with different concentrations of NAA and IBA. Key words: In vitro, Shoot regeneration, Citrus macroptera D.O.I. 10.3329/ptcb.v18i1.3246 Plant Tissue Cult. & Biotech. 18(1): 17-24, 2008 (June)


Author(s):  
N. B. Kravets ◽  
M. Z. Mosula ◽  
N. M. Drobyk ◽  
N. V. Tulaidan ◽  
M. B. Cheterbok

Aim. The aim of the study was to choose conditions for rooting improvement of in vitro cultivated plants of some species of Carlina L. genus. Methods. For receiving and rooting of aseptic sprouts, seeds of Carlina acaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl were subjected to presowing treatment with gibberellic acid solution (GA3) or indolebyturic acid solution (IBA). Sterilized seeds were planted in sterile Petri dishes on semi-solid Murashige, Skoog nutrient medium with half-strength concentrations of macro- and microsalts without growth regulators. Results. It was found that with the seed soaking of C. acaulis, C. cirsioides and C. onopordifolia in GA3 solution the percentage of root formation amounted to 33.3 %, 33.3 % and 22.2 % respectively. Presowing treat-ment of carlina seeds in IBA solution with concentration of 1000 mg for 2–4 hours before sterilization gave a positive effect: the percentage of root formation for C. acaulis, C. cirsioides and C. onorordifolia was 2.4–4.5 times higher compared to the treatment with GA3 solution. Conclusions. To form the root system of carlina plants it is effective to soak the seeds in the solution of IBA. Thus we were able to increase the percentage of rooting of C. sirsioides and C. onorordifolia plants to 100 %, C. acaulis plants – up to 80 % and avoid sprouts’ injury and changes in the concentra-tions of the IBA, which may occur during sterilization at high temperatures by using non-sterile solution of growth regulators. Keywords: Carlina àcaulis L., Carlina cirsioides Klok, Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl, in vitro, sprouts rooting.


1948 ◽  
Vol s3-89 (7) ◽  
pp. 239-252
Author(s):  
P. B. MEDAWAR

The transplantation of skin from one rabbit to another elicits a reaction that conforms in main outline with that of an actively acquired immunity. The experiments described in this paper were designed to test the hypothesis that the regression of such grafts is secured by the action of antibodies demonstrable in vitro. Skin from adult rabbits has therefore been cultivated in the presence of serum and growing mesenchymal tissues derived solely from rabbits heavily and specifically immunized against it. Immune sera and tissues are without effect on the survival, cell-division frequency and migratory activities of explanted skin, and agglutinins for epidermal cell suspensions are not demonstrable in immune sera. With certain stated qualifications, it has therefore been concluded that the occurrence of free antibodies is not a sufficient explanation of the regression of skin homografts in vivo.


2018 ◽  
Vol 28 (2) ◽  
pp. 215-222 ◽  
Author(s):  
Arun Kumar Khajuria ◽  
NS Bisht

An efficient indirect plant regeneration protocol was developed for Viola canescens, an important medicinal herb used in broad spectra of diseases in number of folk medicines since aeon. Excessive use of this plant without any rehabilitating measure has led to decline its natural population. Present investigation reports the use of zeatin to regenerate the plant from the callus on MS following its acclimatization on the soil condition. Calli of the plant responded positively to zeatin and maximum number of shoots 13.07 ± 2.01 were obtained when 9.12 μM concentration of zeatin was used. Regenerated shoots were subsequently rooted with IBA on MS and half strength MS and showed maximum number of roots 14.13 ± 1.64 after 60 days when medium was fortified with 4.92 μM IBA, followed by transferring them to soil condition, acclimatization of the plantlet was carried in growth chamber and then finally to the field for their survival where it showed 80% survival. Plant Tissue Cult. & Biotech. 28(2): 215-222, 2018 (December)


HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.


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