scholarly journals In vitro Regeneration of Alstroemeria cv. ‘Balance’ Based On Direct Organogenesis

2017 ◽  
Vol 14 (2) ◽  
pp. 557-566
Author(s):  
Maryam Beigi Harchegani ◽  
Hossein Nazarian ◽  
Mahmoud Otroshy ◽  
Mohammad Ali Ebrahimi ◽  
Ali Motamedi
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Direct Regeneration ◽  
Direct Organogenesis ◽  
Root Induction ◽  
Factorial Experiments ◽  
Apical Bud

ABSTRACT: The present study aimed at improving the efficiency of the in vitro regeneration of Alstroemeria cv. ‘Balance’ protocol through direct organogenesis technique using three different origins of explants (nodal stem, rhizome apical bud, rhizome segments) as two separate factorial experiments with completely randomized design with six replications which were implemented in three stages. Firstly, Direct regeneration, including two factorial experiments to induce regeneration in organogenesis media by utilizing the four NAA concentrations (0, 0.5, 1, 2 mg/l) combined with five BAP concentrations (0, 0.5, 1, 1.5, 2 mg/l) in the first experiment, and TDZ concentrations (0, 0.5, 10 mg/l) in combination with three IBA concentrations (0, 1, 2 mg/l) in the second experiment. Secondly, shoot regeneration and elongation of stems in regenerated buds in the MS basal medium and finally, rooting of the regenerated shoots in the root induction mediums by NAA and IBA. Results of regeneration experiment revealed that, in the culture medium containing 10 mg/l TDZ in combination with 2 mg/l IBA and rhizome apical bud explants, the maximum rates of regeneration percentage, the highest number and length of the shoots were produced. Resulted shoots produced the greatest number of roots and rhizomes in the culture medium comprising 1 mg/l NAA in combination with 2 mg/l IBA as auxins. From the two tested explants, rhizome apical bud, was known as the best explants for shoot regeneration obtained plantlets successfully adapted to environmental conditions and were transferred to the greenhouse. Results of current study indicated that Alstroemeria can be produced through direct organogenesis.

scholarly journals In-vitro Regeneration from Direct and Indirect Organogenesis of Crescentia alata Kunth an Important Multipurpose Medicinal Tree

2021 ◽  
pp. 48-58
Author(s):  
R. Abinaya
Keyword(s):  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Direct Organogenesis ◽  
Ms Medium ◽  
Indirect Organogenesis ◽  
Medicinal Tree ◽  
Short Period ◽  
In Vitro Clonal Propagation

In this present work, an in-vitro regeneration protocol for Crescentia alata (C. alata) was developed using various explants on Murashige and Skoog (MS) medium augmented with different concentrations and combinations of plant growth regulators (PGRs) for direct and indirect regeneration. The direct organogenesis was established from nodes and internodes on MS medium supplemented with cytokinins and auxins. The indirect organogenesis via callus phase was obtained from leaf, nodes and internodes on MS medium supplemented with different concentrations of PGRs. The high frequency shoot organogenesis were achieved directly from nodal explants were cultured on MS medium supplemented with 3.0 mg/L BAP+0.5 mg/L KIN +1.0 mg/L NAA. Indirect organogenesis callogenic frequency was optimized at the concentration of MS medium containing 1.0 mg/L BAP + 5.0 mg/L IAA. The callus was obtained from all the explants were used, among these explants internodal explants gave best result on MS medium supplemented with different concentrations of cytokinins and auxins for indirect organogenesis experiment. Indirect organogenesis the highest number of shoot regeneration was obtained in MS Basal Medium with 4.0 mg/L BAP + 0.5 mg/L KIN + 2.0 mg/L NAA from internodal explants. For root formation the regenerative shoots which were sub cultured on MS medium containing different ratios of auxins. The rooted plantlets were transferred successfully to the pots containing sterilized soil and were successfully hardened at greenhouse condition for 20 days then exposed to the natural environment. This is the first successful micropropagation report of an efficient and rapid in-vitro clonal propagation protocol for C. alata by direct and indirect shoot organogenesis through various explants, which can be employed for conservation of this important medicinal tree species as well as the utilization of an biologically important active biomolecules. This protocol can be very useful to obtain plants from various explants, without the requirement of meristematic regions, enabling the obtainment of a higher number of plants in short period.

scholarly journals IN VITRO Regeneration of Bina Mungbean Varieties

2015 ◽  
Vol 7 (2) ◽  
pp. 47-52 ◽  
Author(s):  
S Mojumder ◽  
MD Hossain ◽  
MS Haque ◽  
KM Nasiruddin
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Root Length ◽  
In Vitro Regeneration ◽  
Root Induction ◽  
Ms Medium ◽  
Root Initiation ◽  
Root Proliferation ◽  
Callus Initiation

The experiment was conducted to develop an efficient protocol for in vitro regeneration of mungbean (Vignaradiata) on the aspect of regeneration potentiality of two mungbean varieties (BINA mung 5 and BINA mung 7) as influenced by different combinations of growth regulators supplemented with MS medium. Cotyledon explant of both varieties was used for the present study. Data were collected for various characters of callus initiation, shoot regeneration and root proliferation. Initiation of callus (%) and required days for its initiation and weight of callus were influenced significantly due to the effect of varieties where BINA mung 5 produced more callus induction (40.36%) at minimum requiring time (18.27 days) including heavier sizes of callus (1.54 g) than BINA mung 7 when BINA mung 5 further recorded the longest root (2.92 cm) compare to BINA mung 7. Effect of treatments of the present study were significantly influenced the whole characters regarding callus culture, shoot regeneration and root proliferation. The highest percentage of callus (88.44%) within minimum time (12.53 days) including larger sizes callus (3.521 g) were produced in 1.0 mg L–1 BAP + 2.5 mg L–1 NAA among the treatments while the highest percentage of regenerated shoot (83.44%) at minimum requiring time (17.59 days) and more shoots (7.69 callus–1) were obtained in 1.0 mg L–1 BAP + 2.0 mg L– 1 NAA. Root induction (82.50%), number of roots plantlet–1 (8.469) with minimum requiring time for initiation (14.13 days) and root length (5.250 cm) were the highest in 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP. Incase of interaction, percentage of callus initiation (89.38 %) was the highest in BINA mung 5 treated by 1.0 mg L–1 BAP + 2.5 mg L–1 NAA at requiring minimum time (12.38 days) while same treatment produced the larger callus (3.581 g) among the interactions. The highest percentage (84.38%) and number (7.813 callus–1) of shoot with minimum requiring time (17.50 days) were found from BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. Similarly, the longest shoot (5.58 cm) was produced from the BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. However, root induction (%), roots plantlet–1, days required for root initiation and root length were statistically similar among the whole interaction treatments due to non significant variation. This result mentioned that the variety BINA mung 5 was better than BINA mung 7 for callus induction, shoot regeneration and root initiation while 1.0 mg L–1 BAP + 2.5 mg L–1 NAA, 1.0 mg L–1 BAP + 2.0 mg L–1 NAA and 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP supplemented with MS medium were the best combinations for better callusing, higher ability of shoot regeneration and root proliferation.DOI: http://dx.doi.org/10.3329/jesnr.v7i2.22203 J. Environ. Sci. & Natural Resources, 7(2): 47-52 2014

scholarly journals Propagation of Sedum spectabile Boreau in Leaf Culture in Vitro

2012 ◽  
Vol 40 (1) ◽  
pp. 107 ◽  
Author(s):  
Cuiqin YANG ◽  
Yaoguo QIN ◽  
Xin SUN ◽  
Shu YUAN ◽  
Honghui LIN
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
Shoot Proliferation ◽  
Shoot Formation ◽  
Root Induction ◽  
Ms Medium ◽  
Shoot Induction ◽  
Leaf Base ◽  
Stem Segments

An efficient protocol was established for Sedum spectabile Boreau propagation. Various leaf parts were used as explants to regenerate plantlets, the stem segments of which were cultured for shoot proliferation and plantlet multiplication. The results showed that the leaf base was the optimal explant, as compared to both the middle and the top of leaves, for shoot formation. The highest shoot induction of 88.9% was observed on MS medium supplemented with 0.6 mg/l TDZ and 0.1 mg/l NAA. Hyperhydric leaves obtained in primary culture developed first into abnormal somatic embryos 10 days after subculture, and then into hyperhydric plantlets after an additional 10 days. The hyperhydric plantlets reversed to normal plantlets when plant growth regulators were removed from culture medium. Further, stem segments from reversed plantlets were used for shoot regeneration and root induction. Optimal shoot regeneration was obtained in MS medium containing 0.6 mg/l TDZ with 0.1 mg/l NAA. Root induction and root mean number were all higher on auxin-free medium than on medium containing auxins.

scholarly journals In vitro propagation of 'Echo' cultivars of Eustoma grandiflorum (Raf.) Shinn.

2005 ◽  
Vol 11 (2) ◽  
Author(s):  
M. Ördögh ◽  
E. Jámbor-Benczúr ◽  
A. Tilly Mándy
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Propagation ◽  
Culture Medium ◽  
Basal Medium ◽  
The Other ◽  
Eustoma Grandiflorum ◽  
After Effect ◽  
Number Of Shoots

Eustoma grandiflorum (Raf.) Shinn. 'Echo' Fl cultivars ('Echo White', 'Echo Rose', 'Echo Blue', 'Echo Blue Picotee') were used and multiplication of shoots was evaluated on Murashige and Skoog (1962) basal medium with 11 g/1 agar-agar and 20 g/1 sucrose. To test the effect of BA different concentrations were added: 0.10, 0.25 mg/1 and a culture medium without BA. Differentiation of roots was examined on Jámbor-Benczúr and Marta (1990) basal medium with the same concentration of agar-agar and sucrose. To examine the effect on rooting, various concentrations of NAA were used: 0.5, 1.0, 2.0, 3.0 mg/l. The pH was adjusted to 5.6 in every case using KOH. We studied the after-effect of different concentrations of BA during the acclimatisation. During the multiplication, the cultivar 'Echo White' formed the most shoots and the smallest leaves on the medium with 0.10 mg/1 BA. Fortunately, in the case of this cultivar, the number of shoots was reduced and the length of leaves was increased succesfully on the medium without BA. The other three cultivars developed the longest leaves on the medium containing 0.10 mg/1 BA. Sometimes not only shoot regeneration but spontaneous rooting was observed during the multiplication. Examining the rooting, the highest percent of roots was found on the medium with 1.0 mg/1 NAA, and the cultivar 'Echo Rose' formed the most roots on this medium. Higher concentration (2.0 and 3.0 mg/1) of NAA already reduced the number of roots in all of the cultivars. During the acclimatisation, the percentage of survival was 76.3% and the tallest plants with the longest leaves were found on the multiplication medium with 0.25 mg/1 BA. 'Echo Blue Picotee' gave the best results with the tallest pieces and longest leaves on this medium.

scholarly journals In Vitro Adventitious Rooting of Carrizo Citrange Microshoots

HortScience ◽  
2010 ◽  
Vol 45 (6) ◽  
pp. 988-990 ◽  
Author(s):  
Almudena Montoliu ◽  
Aurelio Gómez-Cadenas ◽  
Rosa M. Pérez-Clemente
Keyword(s):  
Acetic Acid ◽  
Gibberellic Acid ◽  
Activated Charcoal ◽  
Culture Medium ◽  
Basal Medium ◽  
Poncirus Trifoliata ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Carrizo Citrange

The objective of this work was to develop an efficient in vitro rooting protocol for one of the most commercially used citrus rootstocks in Spain, Carrizo citrange (Citrus sinensis L. Osbeck × Poncirus trifoliata L. Raf.). Single-node cuttings taken from greenhouse-grown plants were cultured in petri dishes containing basal Murashige and Skoog medium. Shoots from nodal stem segments were excised and cultured in a multiplication medium (basal medium supplemented with 1.8 μM 6-benzylaminopurine) to promote the development of axillary buds. Individual shoots (15 mm long) were treated with different hormones at several concentrations for root induction evaluations. The addition of activated charcoal (AC) to the culture medium was also explored. The addition of auxins to the culture medium enhanced rooting percentage. Optimal results were obtained when 1-naphthalene acetic acid (10.8 μM) and gibberellic acid (0.3 μM) were added to the culture medium. The addition of AC to the rooting medium resulted in negative effects on the percentage of rooted shoots but had a positive effect on number of roots per rooted shoot. Chemical names used: activated charcoal (AC); 6-benzylaminopurine (BA); 1-naphthalene acetic acid (NAA); gibberellic acid (GA3); indole-3-butyric acid (IBA)

scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

2019 ◽  
Vol 29 (1) ◽  
pp. 81-97
Author(s):  
Sujay Kumar Bhajan ◽  
Setara Begum ◽  
Mohammad Nurul Islam ◽  
M Imdadul Hoque ◽  
Rakha Hari Sarker
Keyword(s):  
Genetic Transformation ◽  
Vigna Radiata ◽  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Pcr Analysis ◽  
Root Induction ◽  
Half Strength

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

scholarly journals In vitro Regeneration of Cabbage (Brassica oleracea L. var. Capitata) through Hypocotyl and Cotyledon Culture

1970 ◽  
Vol 17 (2) ◽  
pp. 131-136 ◽  
Author(s):  
M.K. Munshi ◽  
P.K. Roy ◽  
M.H. Kabir ◽  
G. Ahmed
Keyword(s):  
Shoot Regeneration ◽  
Callus Formation ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Direct Regeneration ◽  
Cotyledon Explants ◽  
Cotyledon Culture ◽  
Best Response ◽  
Half Strength

The best response toward direct regeneration of multiple shoots from seven-dayold seedling was observed on half-strength MS supplemented with 0.5 mg/l BA. Hypocotyl and cotyledon explants produced highest percentage (73 and 66, respectively) of shoots. The maximum number of shoots (12) and the highest shoot length of 5.9 cm were also observed in this medium. On the other hand, indirect regeneration via callus was observed in cotyledonary explants. Maximum percentage of callus formation was observed on MS containing 1.0 mg/l 2,4-D and 0.5 mg/l NAA. Highest frequency of shoot regeneration was achieved on MS fortified with 2.0 mg/l BA and 0.1 mg/l NAA in cotyledon derived callus. Shoot regeneration was not obtained in hypocotyl-derived callus. Shoots rooted well when they were excised individually and implanted in halfstrength MS with 0.5 mg/l IBA in which 98% rooting was achieved within 10 - 12 days. The well rooted in vitro raised plantlets were successfully transferred to soil and their survival rate under natural environment was 86%. Key words: In vitro regeneration, Cabbage, Hypocotyl, Cotyledon  D.O.I. 10.3329/ptcb.v17i2.3233  Plant Tissue Cult. & Biotech. 17(2): 131-136, 2007 (December)

scholarly journals IN VITRO REGENERATION OF POTATO (Solanum tuberosum L.)

2011 ◽  
Vol 24 (1) ◽  
pp. 07-14
Author(s):  
M. M. H. Molla ◽  
K. M. Nasiruddin ◽  
M. Al-Amin ◽  
M. S. Haque ◽  
D. Khanam
Keyword(s):  
Agricultural Research ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Zeatin Riboside ◽  
Direct Regeneration ◽  
Step Procedure ◽  
In Vitro Shoots ◽  
Nodal Cuttings

The experiment was conducted during the period from January to December, 2008 at the Laboratory of Biotechnology, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh. A two step procedure was followed for direct plant regeneration of potato. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l-1 IBA and 30 g sugar was used for in vitro potato plants development via nodal cuttings. Leaves and internodes of in vitro grown potato variety Asterix was cultured in instant MS basal medium supplemented with 0.01 mg l-1 IAA, 0.20 mg l-1 GA3, 44.0 g l-1 CaCl2, 20 g sucrose. Eight different concentrations of Zeatin riboside (ZR) viz., 0, 0.1, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 mg l-1 along with above supplements (step II) were tested for in vitro direct regeneration of potato. MS medium supplemented with 4-5 mg l-1 of ZR performed better in respect of shoot induction from internodal explants. All the internodes produced shoots directly from the basal and apex region within 18-21 days and from the leaves within 21-28 days in 2 - 5 mg l-1 ZR. Maximum 43.20 and 7.25 shoots were recorded from each internode and leaf explant, respectively. In vitro shoots treated with 0.5 mg l-1 of IBA produced roots profusely within 21 days.DOI: http://dx.doi.org/10.3329/bjpbg.v24i1.16307

scholarly journals In vitro micropropagation of Jasminum grandiflorum L.

2018 ◽  
Vol 53 (4) ◽  
pp. 277-282
Author(s):  
Md S Rahman ◽  
NJ Mouri ◽  
NC Nandi ◽  
S Akter ◽  
Md S Khan
Keyword(s):  
Culture Medium ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Soil Condition ◽  
Coconut Water ◽  
Garden Soil ◽  
In Vitro Micropropagation ◽  
Axillary Meristems

An efficient protocol was developed for in vitro plant regeneration of Jasminum grandiflorum L. The highest elongated shoots (60%) were achieved from axillary meristems using MS (Murashige and Skoog ) basal medium supplemented with 1 mg/l 6-benzyladenine (BAP) and 60 mg/l coconut water. After adding 1.0 mg/l BAP and 45 mg/l coconut water in the culture medium, the highest rate of shoot proliferation was exhibited after 4 weeks of culture. Rooting was found within 14-23 days after the cut end of shoots was soaked in 1.5 mg/l IBA solution for 3-7 minutes. The regenerated healthy rooted plantlets were transferred to small plastic pot containing garden soil and compost in a ratio of 2:1. Maximum (75%) in vitro rooted plants were survived in the shade house and finally survived naturally in soil condition. The successful protocol for in vitro regeneration was developed which will facilitate the conservation and propagation of the important medicinal plant.Bangladesh J. Sci. Ind. Res.53(4), 277-282, 2018

An effective protocol for improved regeneration capacity of Kabuli chickpeas

2012 ◽  
Vol 92 (6) ◽  
pp. 1057-1064 ◽  
Author(s):  
I. S. Yadav ◽  
N. P. Singh
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
Genetic Manipulation ◽  
In Vitro Regeneration ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Regeneration System ◽  
Embryonic Axes

Yadav, I. S. and Singh, N. P. 2012. An effective protocol for improved regeneration capacity of Kabuli chickpeas. Can. J. Plant Sci. 92: 1057–1064. An efficient protocol for in vitro regeneration is essential for genetic manipulation and micro-propagation of important plant species. A direct shoot regeneration system has been optimized for Desi chickpeas, but an effective regeneration protocol is still needed for Kabuli chickpeas. An efficient regeneration protocol for Kabuli chickpeas was developed, using whole embryonic axes, an embryonic axes slice and cotyledonary node explants from two genotypes L550 and JGK-1. Depending upon chickpea genotype, type of explant and culture medium, percentage of shoot producing explants (frequency) and the number of shoots per explant (efficiency) varied from 10 to 83% and from 1 to 58, respectively. The shoot regeneration capacity (SRC=frequency×efficiency), which is an indicator of the effectiveness of the protocol, varied from 47 to 2508 shoots per 100 explants cultured. On average, SRC of L550 was 1.8 times higher than JGK-1. Murashige and Skoog's (MS) medium+B5 vitamins supplemented with 8.0 µM benzyl amino purine (BAP)+0.5 µM α- naphthalene acetic acid (NAA) and 0.1 M sucrose plus embryonic axes was found to be the most effective culture medium and type of explants, respectively. Half strength MS medium+2% sucrose supplemented with 4 µM NAA, 3µ M IAA or 4µM IAA produced a high rooting percentage in both chickpea genotypes. The regeneration process starting from explant preparation to establishment of a complete plant in soil took 105–110 d. This optimized regeneration method holds promise for facilitating the insertion of interested genes through genetic transformation for improvement of Kabuli chickpeas.

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