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.

scholarly journals Efficient Regeneration of Tobacco (Nicotiana tabacum L.) Plantlets from Cotyledon, Hypocotyl and Leaf Explants: An Excellent Model Plant for Gene Function Analysis

2020 ◽  
pp. 1-9
Author(s):  
Md. Shoyeb ◽  
Kanis Fatema ◽  
Md. Abdur Rauf Sarkar ◽  
Atikur Rahman ◽  
Shaikh Mizanur Rahman
Keyword(s):  
Gene Function ◽  
Callus Induction ◽  
Function Analysis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Regeneration Ability ◽  
Ms Medium ◽  
Gene Function Analysis

Tobacco has been widely used as a model plant for stable and non-stable gene function analysis. Successful Agrobacterium-mediated transformation mainly depends on in vitro regeneration of tobacco plant. However, a reliable and standard regeneration protocol of tobacco using multiple explants is limited. In this study, we established a reliable and reproducible regeneration protocol of tobacco using three different explants i.e. cotyledon, hypocotyl and leaf. Preliminary, surface sterilized tobacco seeds were germinated on growth regulator free MS medium. Thereafter, in vitro germinated explants were inoculated into Murashige and Skoog [1] media supplemented with different combination and types of growth regulators for callus induction and subsequent regeneration of plantlets. It was revealed that, regeneration ability of explants is greatly influenced by type and nature of the explant. Among the three explants, higher callus induction (95%) was obtained in MS medium supplemented with 2.0 mg l-1 kinetin + 2.0 mg l-1 IAA from leaf explant. Also, leaf explant exhibited much higher regeneration ability (95%) than hypocotyl (60%) and cotyledon (45%) explants. Significantly highest number of shoots (8.0) were regenerated from leaf explants cultured on MS medium supplemented with 3.0 mg l-1 Kinetin+1.0 mg l-1 IAA compared to the other hormone combinations. Regenerated mature shoots were showed normal root after transferred onto ½ MS medium containing 0.3 mg l-1 IBA. This study will provide valuable information related to in vitro regeneration of tobacco plantlets using cotyledon, hypocotyl and leaf explants and will be used as a standard protocol for Agrobacterium-mediated transformation for gene function analysis.

scholarly journals In vitro Micropropagation of Abutilon indicum L. through Leaf Explants

1970 ◽  
Vol 19 (2) ◽  
pp. 177-184 ◽  
Author(s):  
Jyoti Ranjan Rout ◽  
Manorama Mishra ◽  
Ritarani Das ◽  
Santi Lata Sahoo
Keyword(s):  
Callus Induction ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Plantlet Regeneration ◽  
Root Induction ◽  
Ex Vitro ◽  
In Vitro Micropropagation ◽  
Half Strength ◽  
Abutilon Indicum

The present investigation was conducted to develop a protocol for rapid callus induction and plant regeneration from leaf explant of Abutilon indicum L. Callus induction and plantlet regeneration at various frequencies were observed on MS using different concentrations of 2,4-D alone or in combination with BAP and Kn. The highest percentage of callus induction was observed with 2.5 mg/l 2,4-D (90) and with combination of 0.5 mg/l Kn (85). Optimum shoot formation was observed on same medium but supplemented with 2.0 mg/l Kn and 1.0 mg/l NAA (11.2). Rooting experiments with half strength of MS revealed that NAA was more suitable for root induction compared to IBA and IAA. The healthy in vitro rooting plantlets were successfully transferred to the field. The survival of the plantlets under ex vitro condition was 87%. Key words: Abutilon indicum, Callus induction, Leaf explants, Micropropagation D.O.I. 10.3329/ptcb.v19i2.5435 Plant Tissue Cult. & Biotech. 19(2): 177-184, 2009 (December)

scholarly journals INDUCTION OF CALLOGENESIS AND SHOOT REGENERATION OF A MEDICINAL PLANT SPECIES PERISTROPHE BICALYCULATA (RETZ.) NEES.

2015 ◽  
Vol 2 (2) ◽  
pp. 55-59
Author(s):  
Femila Jose V ◽  
Asir Selin Kumar R
Keyword(s):  
Medicinal Plant ◽  
Plant Species ◽  
Callus Induction ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Medicinal Plant Species ◽  
Multiple Shooting ◽  
Ms Medium ◽  
Apical Leaf ◽  
Peristrophe Bicalyculata

In the present study, protocol for callus induction and regeneration for the medicinal plant species, Peristrophe bicalyculata (Retz.)Nees has been developed by using leaf explants. Young apical leaf explant was used for callus induction on MS medium containing BAP and NAA at 2`0 and 0.8 mgl-1 respectively showedmaximum callus induction (80%). The amount of callus responded for shoot formation (81%) was obtained in the MS medium containing BAP (2.0 mgl-1) and GA3 (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (2.0 mgl-1) and IAA (0.2 mgl-1) for shoots rooted. Regeneratedplantlet were successfully acclimatized and hardened off inside the culture and then transferred to green Peristrophe bicalyculata, MS medium, multiple shooting, Acclimatizationhouse with better survival rate

scholarly journals Optimizing the concentrations of plant growth regulators for in vitro shoot cultures, callus induction and shoot regeneration from calluses of grapes

OENO One ◽  
2015 ◽  
Vol 49 (1) ◽  
pp. 37 ◽  
Author(s):  
Nadra Khan ◽  
Maqsood Ahmed ◽  
Ishfaq Hafiz ◽  
Nadeem Abbasi ◽  
Shaghef Ejaz ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
Growth Regulators ◽  
Callus Induction ◽  
Leaf Explants ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Grape Cultivar ◽  
Half Strength ◽  
Number Of Shoots

<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>

scholarly journals In Vitro Regeneration of Dendrobium sp. of Orchid Using Leaf Tip as Explant

2016 ◽  
Vol 8 (2) ◽  
pp. 75-78
Author(s):  
K Goswami ◽  
S Yasmin ◽  
KM Nasiruddin ◽  
F Khatun ◽  
J Akte
Keyword(s):  
In Vitro Regeneration ◽  
Plantlet Regeneration ◽  
Shoot Length ◽  
Ms Medium ◽  
Fresh Weight ◽  
Half Strength ◽  
Maximum Root Length ◽  
Maximum Root ◽  
Regenerated Plantlets

Plant growth regulators (PGRs) namely, 2,4-D, NAA and BAP were added into Murashige and Skoog (MS) medium to observe the effect of PGRs on the growth and development of Dendrobium sp. orchid. Leaf tips of Dendrobium sp. were used as explants and inoculated on MS medium supplemented with 2, 4 D (0, 0.5, 2.5, 5, 10 mgL?1) for development of PLBs. The maximum PLBs formation (90%) and the maximum number of PLBs (16.00) were observed in 10 mgL?1 2, 4-D into MS medium after 60 days of culture. Subcultured PLBs were inoculated on MS medium supplemented with different combinations of NAA (0, 0.5, 2.5, 5 mgL?1) and BAP (0, 0.5, 2.5, 5 mgL?1) for shoot regeneration. The maximum number of shoot (11.00), the highest fresh weight (0.6233g) and the highest shoot length (3.613 cm) were observed in 0.5 mgL?1 NAA + 0.5 mgL?1 BAP after 60 days of culture. Even, the maximum number of root (4.00), the maximum root length (1.627cm) and the maximum plantlet regeneration percentage (93.33%) were observed with the combined effect of 0.5 mg NAA and 0.5 mg BAP after 60 days of culture. Finally, regenerated plantlets were transferred into half strength MS medium to obtain plants.J. Environ. Sci. & Natural Resources, 8(2): 75-78 2015

scholarly journals In vitro regeneration of popular tobacco varieties of Bangladesh from leaf disc

1970 ◽  
Vol 35 (1) ◽  
pp. 125-134 ◽  
Author(s):  
MA Rahman ◽  
MA Alam ◽  
MR Hossain ◽  
A Hossain ◽  
R Afroz
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
In Vitro Regeneration ◽  
Leaf Disc ◽  
Shoot Formation ◽  
Plantlet Regeneration ◽  
Regeneration Ability ◽  
Ms Medium ◽  
Leaf Discs

Regeneration ability of five Nicotiana varieties viz., Virginia, Jati, Motihari, CC Bengal and Sumatra were investigated via callus induction using leaf discs. Explants were cultured on MS medium supplemented with different concentrations and combinations of plant growth regulators. Callus formation frequency was 67.20%. Among the varieties used, Motihari induced the highest percentage (97.50%) of callus followed by Jati (92.50%) in 2.0 rng/L Kinetin and 2.0 mg/L IAA. Shoots were induced from calli cultured on the same medium. Maximum shoot formation from leaf discs was 82.50% on medium supplemented with 2.0 mg/L Kinetin and 2.0 mg/L IAA. It was also revealed from this study that Motihari was the best variety for callus formation and subsequent plantlet regeneration which is a pre-requisite for vector mediated transformation for varietal improvement of Nicotiana species. The rooting response of regenerated shoots was observed by using ½ MS medium with IBA (0.0, 0.5, and 1.0 mg/L). The highest root formation was found in Motihari (90%) with ½ MS medium supplemented with 0.5 mg/L IBA. After that regenerated plantlets with plenty of roots were transferred successfully to pots and subsequently to the field. Keywords: Tobacco; Nicotiana; in vitro regeneration; callus induction; plantlet regeneration; leaf disc; phytohormone. DOI: 10.3329/bjar.v35i1.5873Bangladesh J. Agril. Res. 35(1) : 125-134, March 2010

scholarly journals An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501
Author(s):  
Sanjog T. Thul ◽  
Arun K. Kukreja
Keyword(s):  
Shoot Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Ms Medium ◽  
Mentha X Piperita ◽  
Half Strength ◽  
Multiple Shoot Regeneration

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

scholarly journals In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Genetic Transformation ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Native Forest ◽  
Ms Medium ◽  
Dalbergia Sissoo ◽  
Callus Induction Medium

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

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 Callus Induction in Baru (Dipteryx alata Vog.) Explants

2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

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