scholarly journals An Improved System for Rapid in vitro Regeneration of Saintpaulia ionantha

2014 ◽  
Vol 24 (1) ◽  
pp. 37-45
Author(s):  
Zahra Ghorbanzade ◽  
Mohammad Ahmadabadi
Keyword(s):  
Root Formation ◽  
High Efficiency ◽  
Genetic Manipulation ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Time Saving ◽  
Rapid Multiplication ◽  
Saintpaulia Ionantha ◽  
Regenerated Plantlets

For rapid multiplication and genetic manipulation of African violets (Saintpaulia ionantha Wendl.) was aimed at developing a rapid and efficient regeneration and adaptation system from leaf explants. Using RM medium supplemented with different combinations of growth regulators, we developed a highly efficient and time-saving in vitro regeneration protocol. The developed system was also successfully applied for plant regeneration from petiole and internode explants of several Saintpaulia cultivars at a high efficiency. Regeneration occurred through both somatic embryogenesis and shoot organogenesis at a high frequency. Also, we developed a rapid root formation protocol followed by their efficient adaptation for in vitro regenerated plantlets. Plant Tissue Cult. & Biotech. 24(1): 37-45, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19194

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

In vitro differentiation and plant regeneration from root and other explants of juvenile origin in pea (Pisum sativum L.)

2017 ◽  
Author(s):  
Shikha Sharma ◽  
Geetika Gambhir ◽  
D. K. Srivastava
Keyword(s):  
Pisum Sativum ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Cotyledonary Node ◽  
Shoot Elongation ◽  
Root Regeneration ◽  
Pisum Sativum L ◽  
Regenerated Plantlets

In vitro regeneration of pea explants (Pisum sativum L. var. ‘Lincon’) was done in 49 different combinations and concentrations of BAP, BAP and NAA, BAP and IBA, TDZ, TDZ and Adenine for shoot regeneration from hypocotyl, root, leaf and cotyledonary node. High frequency shoot regeneration was obtained in hypocotyl (81.43%), root(83.53%) and cotyledonary node(72.76%) on MS medium supplemented with 4.50 mg/l BAP and 1.86mg/l NAA, 2.00mg/l TDZ and4.50 mg/l BAP and 1.86mg/l NAA respectively. No shoot regeneration was obtained from leaf explants on any of the combination used. Shoot elongation was observed on the same medium used for shoot regeneration respectively.MS medium supplemented with 0.20 mg/l IBA was found best for root regeneration from in vitro raised shoots. The plantlets were able to regenerate within 6-7 weeks. The regenerated plantlets were acclimatized in pre-sterilized cocopeat.

scholarly journals High-Frequency Plant Regeneration, Genetic Uniformity, and Flow Cytometric Analysis of Regenerants in Rutachalepensis L.

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2820
Author(s):  
Ahmed A. Qahtan ◽  
Mohamad Faisal ◽  
Abdulrahman A. Alatar ◽  
Eslam M. Abdel-Salam
Keyword(s):  
Genetic Manipulation ◽  
In Vitro Regeneration ◽  
Carbon Sources ◽  
Flow Cytometric Analysis ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Bioactive Constituents ◽  
Flow Cytometric ◽  
Regenerated Plantlets

Ruta chalepensis L., an evergreen shrub in the citrus family, is well-known around the world for its essential oils and variety of bioactivities, indicating its potential medicinal applications. In this study, we investigated the effect of different culture conditions, including plant growth regulators, media types, pH of the medium, and carbon sources, on in vitro regeneration from nodal explants of R. chalepensis. Following 8 weeks of culture, the highest percentage of regeneration (96.3%) and maximum number of shoots (40.3 shoot/explant) with a length of 4.8 cm were obtained with Murashige and Skoog (MS) medium at pH 5.8, supplemented with 3.0% sucrose and 5.0 µM 6-Benzyladenine (BA) in combination with 1.0 µM 1-naphthaleneacetic acid (NAA). For rooting, individually harvested shootlets were transferred on ½ MS (half-strength) supplemented with IAA (indole-3-acetic acid), IBA (indole 3-butyric acid), or NAA, and the best response in terms of root induction (91.6%), number of roots (5.3), and root mean length (4.9 cm) was achieved with 0.5 µM IBA after 6 weeks. An average of 95.2 percent of healthy, in vitro regenerated plantlets survived after being transplanted into potting soil, indicating that they were effectively hardened. DNA assays (PCR-based markers) such as random amplification of polymorphic DNA (RAPD) and directed amplification of minisatellite-region (DAMD) were employed to assess in vitro cultivated R. chalepensis plantlets that produced a monomorphic banding pattern confirming the genetic stability. Additionally, no changes in the flow cytometric profile of ploidy between regenerated plantlets and donor plants were detected. Regeneration of this valuable medicinal plant in vitro will open up new avenues in pharmaceutical biotechnology by providing an unconventional steadfast system for mass multiplication and might be effectively used in genetic manipulation for enhanced bioactive constituents.

scholarly journals In vitro Regeneration of Wedelia calendulacea Less. - A Valuable Endangered Medicinal Plant

2020 ◽  
Vol 30 (2) ◽  
pp. 277-283
Author(s):  
Sabina Yesmin
Keyword(s):  
Root Formation ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Nodal Segments ◽  
Best Response ◽  
Endangered Medicinal Plant ◽  
Regenerated Plantlets

Direct plant regeneration from shoot tips and nodal explants of Wedelia calendulacea Less. was carried out in MS with different concentrations and combinations of BAP, Kn and NAA. Of the two explants nodal segments showed the best response towards the formation of multiple shoots in MS containing BAP (2.0 mg/l), Kn (1.0 mg/l) and NAA (0.5 mg/l). In this combination, the maximum number of shoots/explant obtained was 18.33 ± 0.72 from shoot tips and 26.64 ± 0.47 from nodal explants. Cent per cent root formation was obtained in MS with IBA (0.2 mg/l) within three weeks. The in vitro regenerated plantlets were successfully transplanted to soil for further growth and development. Plant Tissue Cult. & Biotech. 30(2): 277-283, 2020 (December)

scholarly journals High Efficiency In vitro Regeneration and Genetic Stability of Corallocarpus epigaeus - An Endangered Medicinal Plant

2020 ◽  
Vol 30 (2) ◽  
pp. 219-229
Author(s):  
Spoorthi Veera ◽  
Pavani Chirumamilla ◽  
Shasthree Taduri
Keyword(s):  
High Efficiency ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Inter Simple Sequence Repeat ◽  
Multiple Shoot ◽  
Nodal Explants ◽  
Multiple Shoot Induction ◽  
Endangered Medicinal Plant

Indirect regeneration of plantlets from multiple shoot induction of Corallocarpus epigaeus was obtained from leaf and nodal explants on MS with different concentrations of BAP in combination with IAA/IBA or IBA alone. Among all the combinations, BAP and IBA exhibited maximum regeneration. High frequency of multiple shoots (89%) was obtained on BAP (2.0 mg/l) and IBA (1.5 mg/l) in nodal explants. Maximum mean shoot length of 6.8 ± 0.33 cm was obtained in nodal explants cultured on BAP (1.0 mg/l) + IBA (0.5 mg/l), followed by leaf explants with 6.7 ± 0.47 cm on BAP (3.0 mg/l) + IAA (2.5 mg/l). The highest frequency of rooting (88.3%) was obtained on NAA (1.0 mg/l) and IBA (2.0 mg/l) with 21.83 ± 0.57 mean number of roots. The well-rooted healthy plantlets were acclimatized with a survival rate of 80%. Inter simple sequence repeat (ISSR) analysis revealed the genetic similarity of in vitro raised plants with the mother plant. Plant Tissue Cult. & Biotech. 30(2): 219-229, 2020 (December)

scholarly journals Rapid target validation in a Cas9-inducible hiPSC derived kidney model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yasaman Shamshirgaran ◽  
Anna Jonebring ◽  
Anna Svensson ◽  
Isabelle Leefa ◽  
Mohammad Bohlooly-Y ◽  
...  
Keyword(s):  
High Efficiency ◽  
Disease Modeling ◽  
Genetic Manipulation ◽  
Disease Model ◽  
Genetically Engineered ◽  
Model Systems ◽  
Target Validation ◽  
Direct Differentiation ◽  
Kidney Organoids

AbstractRecent advances in induced pluripotent stem cells (iPSCs), genome editing technologies and 3D organoid model systems highlight opportunities to develop new in vitro human disease models to serve drug discovery programs. An ideal disease model would accurately recapitulate the relevant disease phenotype and provide a scalable platform for drug and genetic screening studies. Kidney organoids offer a high cellular complexity that may provide greater insights than conventional single-cell type cell culture models. However, genetic manipulation of the kidney organoids requires prior generation of genetically modified clonal lines, which is a time and labor consuming procedure. Here, we present a methodology for direct differentiation of the CRISPR-targeted cell pools, using a doxycycline-inducible Cas9 expressing hiPSC line for high efficiency editing to eliminate the laborious clonal line generation steps. We demonstrate the versatile use of genetically engineered kidney organoids by targeting the autosomal dominant polycystic kidney disease (ADPKD) genes: PKD1 and PKD2. Direct differentiation of the respective knockout pool populations into kidney organoids resulted in the formation of cyst-like structures in the tubular compartment. Our findings demonstrated that we can achieve > 80% editing efficiency in the iPSC pool population which resulted in a reliable 3D organoid model of ADPKD. The described methodology may provide a platform for rapid target validation in the context of disease modeling.

In vitro regeneration of Persian poppy (Papaver bracteatum)

Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Sara Rostampour ◽  
Haleh Sohi ◽  
Ali Dehestani
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
High Performance ◽  
High Efficiency ◽  
In Vitro Regeneration ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Papaver Bracteatum ◽  
Callus Proliferation

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.

scholarly journals In vitro regeneration performance of Corchorus olitorius

1970 ◽  
Vol 8 (1) ◽  
pp. 1-6 ◽  
Author(s):  
M Hoque ◽  
KM Nasiruddin ◽  
GKMN Haque ◽  
GC Biswas
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Root Formation ◽  
Plantlet Regeneration ◽  
Ms Medium ◽  
Corchorus Olitorius ◽  
The Media ◽  
Regenerated Plantlets

The experiment was conducted during May to December 2008 in the Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh to observe the callus induction, regeneration potentiality and to establish a suitable in vitro plantlet regeneration protocol of Corchorus olitorius. MS medium supplemented with different phytohormone concentrations and combinations were used to observe the callus induction, shoot regeneration and root formation ability of the cotyledon with attached petiole derived explant of three genotypes viz. O-9897, O-72 and OM-1. The highest callus induction (92.85%) was observed in O-9897 followed by O-72 (82.14%) in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype O-9897 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (83.33%) followed by O-72 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on halfstrength of MS media supplemented with 0.6 mg/L IBA in genotype O-9897 (45.00%). The in vitro regenerated plantlets from the genotypes O-9897 could be established in the field. Therefore, the genotypes O-9897 of C. olitorius in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA could be used for callus induction and shoot regeneration. Keywords: Regeneration; Phytohormone; Corchorus olitorius DOI: 10.3329/jbau.v8i1.6390J. Bangladesh Agril. Univ. 8(1): 1-6, 2010

scholarly journals In vitro Regeneration and Rapid Multiplication of Dendrobium bensoniae, an Indigenous Ornamental Orchid

2017 ◽  
Vol 14 (2) ◽  
pp. 24-31 ◽  
Author(s):  
S S Riva ◽  
A Islam ◽  
M E Hoque
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Ms Medium ◽  
Shoot Induction ◽  
Rapid Multiplication ◽  
Number Of Shoots

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration.  It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium.  For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31

scholarly journals In vitro regeneration protocol of Rauvolfia serpentina L.

2018 ◽  
Vol 53 (2) ◽  
pp. 133-138 ◽  
Author(s):  
S Khan ◽  
TA Banu ◽  
S Akter ◽  
B Goswami ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Root Induction ◽  
Ms Medium ◽  
Regeneration System ◽  
Rauvolfia Serpentina ◽  
Number Of Shoots

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13∙4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Bangladesh J. Sci. Ind. Res.53(2), 133-138, 2018

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