scholarly journals Callogenesis in leaves of Kalanchoe pinnata Lam. by 2,4-D and BA action

2014 ◽  
Vol 16 (3 suppl 1) ◽  
pp. 760-764 ◽  
Author(s):  
M.R.A. Santos ◽  
M.G.R. Ferreira ◽  
M.C.M. Guimarães ◽  
R.A. Lima ◽  
C.L.L.G. Oliveira
Keyword(s):  
Callus Induction ◽  
Large Scale ◽  
Leaf Explants ◽  
Ms Medium ◽  
Main Technique ◽  
Kalanchoe Pinnata ◽  
Medicinal Properties ◽  
Industrial Level ◽  
Inflammatory Effect

The Kalanchoe pinnata Lam. is a bush species of the Crassulaceae that is distinguished by its important medicinal properties. Its leaves are used as cataplasm to treat headaches and wounds. There is evidence for a hypotensive and anti-inflammatory effect. Techniques of plant tissue culture have been applied to plant species that produce substances likely to be explored in pharmacology, cell suspension being the main technique. At the industrial level, this method utilizes bioreactors in order to produce secondary metabolites on a large scale. The objective of this study was to evaluate the effects of in vitro combinations of 2,4-dichlorophenoxiacetic acid (2,4-D) and benzylaminopurine (BA) on callus induction in leaf explants of K. pinnata. Leaf fragments were inoculated in MS medium supplemented with 3.0% sucrose, 0.8% agar and factorial combinations of 2,4-D (0.00, 4.52, 9.06, 18.12 µM) and BA (0.00, 4.44, 8.88, 17.76 µM). The cultures were kept in the darkness at 24±2ºC for 50 days. The percentage of callus induction and the area of explants covered by callus cells were evaluated. In the absence of growth regulators, callus induction did not occur, with necrosis of all explants. The highest percentage of callus induction was 100%, obtained with the combination of 9.06 µM 2,4-D and 8.88 µM BA, but the calluses covered only 25% of the leaf area. The most efficient combination was 4.52 µM 2,4-D and 8.88 µM BA, resulting in 91% callus induction with 50 to 100% of the explants being covered by callus cells.

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 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.

scholarly journals In vitro callus induction of gerbera from leaf explant

2020 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Sadia Afrin Jui ◽  
Md. Mijanur Rahman Rajib ◽  
M. Mofazzal Hossain ◽  
Sharmila Rani Mallik ◽  
Iffat Jahan Nur ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Callus Induction ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Indole 3 Acetic Acid

The experiment was designed to evaluate the effect of growth regulators on leaf explant of Gerbera for callus induction. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA) were used to initiate cultures. These were added to Murashige and Skoog medium in different combinations and concentrations. Leaf explants cultured on MS medium supplemented with BAP+ 2, 4-D+ IAA in T4 treatment & BAP+ 2,4-D in T5 treatment showed the best results for callus induction. On the other hand callus was induced early in the combination of BA+ 2,4-D + IAA hormone in T5, T9 & T8 treatment respectively. The rate of callus induction was very low in BA + NAA combinations but it was much earlier.   

scholarly journals Effect of phytohormones on shoot apex and leaf explants of Withania somnifera (Ashwagandha)

2016 ◽  
Vol 8 (1) ◽  
pp. 412-415 ◽  
Author(s):  
Archana Rani ◽  
M. Kumar ◽  
Sanjeev Kumar
Keyword(s):  
Acetic Acid ◽  
Shoot Apex ◽  
Callus Induction ◽  
Withania Somnifera ◽  
Leaf Explants ◽  
Ms Medium ◽  
Mass Propagation ◽  
Best Response ◽  
Dichlorophenoxy Acetic Acid

An efficient protocol for callus induction of Withania somnifera through in vitro culture of shoot apex and leaf explant was standardized. Of the various combinations of phytohormones evaluated, MS media supplemented with 6-furfuryl aminopurine (KIN) 0.5 mg/l + 2,4-dichlorophenoxy acetic acid (2, 4-D) 2.0 mg/l was found to be bestfor mean callus induction (86%) in leaf explants after 6 weeks of culture and in case of shoot apex expant the best response and growth of callusing was observed on MS medium supplemented with 2,4-D 1.0 mg/l + BAP 2.0 mg/l (77%).The response of callus growth increases gradually with the reductions in concentration of KIN in culturemedium of both the explants. This protocol might be used in further research for mass propagation of W. somnifera via indirect regeneration methods.

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 Callus Induction and Organogenesis in Sugarcane (Saccharum officinarum L.) var 93v297

2015 ◽  
Vol 48 ◽  
pp. 14-22
Author(s):  
Arjun ◽  
Rao Srinath
Keyword(s):  
Sugar Cane ◽  
Callus Induction ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Saccharum Officinarum ◽  
Ms Medium ◽  
Sand Soil ◽  
Maximum Root Length ◽  
Maximum Root

An efficient protocol for induction of callus and regeneration of a sugar cane var 93v297 has been developed and reported here. Callus induction from immature young leaf explants derived from 2-3-month-old plants was achieved on Murashige and Skoog’s (MS) medium supplemented with different auxins viz, 2,4-D, NAA and IAA. Among different auxins, 2, 4-D at 3.5mg/l + 0.5mg/l BAP was found favourable in inducing callus. Addition of coconut milk and BAP further enhanced the growth of callus maximum being on MS medium supplemented with 0.5mg/l BAP (3602.33±0.88mg). Calli were further evaluated for regeneration. MS medium supplemented with 1.0 mg/l BAP was found suitable where 100% calli regenerated with maximum number of multiple shoots per callus mass (41.40±0.89). Highest number of root emergence (28.33±1.16) and maximum root length (3.40±0.67cm) was achieved on MS medium supplemented with 3mgl/l NAA. The in vitro grown plants were transferred to polycups containing a mixture of sterilized sand, soil and cocopeet (1:1:1) for hardening. The hardened plants were transferred to green-house conditions where they survived with 90% frequency.

scholarly journals In vitro cultivation and regeneration of Solanum melongena (L.) using stem, root and leaf explants

1970 ◽  
Vol 1 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Bishnu Pada Ray ◽  
Lutful Hassan ◽  
Smreeti Kana Sarker
Keyword(s):  
Key Words ◽  
Callus Induction ◽  
Callus Formation ◽  
Solanum Melongena ◽  
Leaf Explants ◽  
In Vitro Cultivation ◽  
Ms Medium

The treatment combinations was BAP (0, 2.0, 3.0 and 4.0 mg/L) and NAA (0, 0.1, 0.5, and 1.0 mg/L). The rate of callus formation varied in different treatments. The highest amount of callus (48.66%) was produced on MS medium containing 2.0 mg/l BAP and 0.5 mg/l NAA from stem and 8.2 days required for callus induction. The number of shoot regenerated through callus from stem containing 2.0 mg/l BAP and 0.5 mg/l NAA was 3.4 (23.287%) and days required for 38.8 days. Key words: Regeneration; BAP; NAA. Nepal Journal of Biotechnology. Jan. 2011, Vol. 1, No. 1 : 49-54

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 Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

2022 ◽  
Vol 0 (0) ◽  
Author(s):  
Hamid Reza SABAGHI ◽  
Gholamreza SHARIFI-SIRCHI ◽  
Pejman AZADI ◽  
Mohammad Hossein AZIMI
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Dianthus Caryophyllus ◽  
Casein Hydrolysate ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Rate ◽  
Shoot Number

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

scholarly journals Callus Induction and Phytochemical Constituents of Finger Eggplant (Solanum sp.)

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Norizzah Jaafar SIDIK ◽  
Norhayati DAUD ◽  
Som Cit SINANG ◽  
Nurul Fazira OMAR
Keyword(s):  
Callus Induction ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Fresh Weight ◽  
Leaf Extracts ◽  
In Vitro Plantlets ◽  
Phytochemical Constituents

This study examined the efficiency of callus induction on optimum concentrations of NAA (a-naphthaleneacetic acid) and BAP (6-benzyladenine) from culturing stem and leaf explants of finger eggplant (Solanum sp.) and investigated the phytochemical constituents of callus tissue. Seeds were sterilized by using 3 and 5 % Clorox solution, which gave the highest number of survival seeds (100 %) and were grown in vitro plantlets. The highest frequency of callus induction (100.00 ± 0.00 %) was obtained from stems and leaf explants that were excised from in vitro plantlets. The stem explants cultured on MS medium consisted of 1.0 mg/L NAA + 1.0 mg/L BAP, giving the maximum mean callus fresh weight (0.14 ± 0.05 g). Meanwhile, the leaf explants cultured on MS medium consisted of  0.5 mg/L NAA + 2.0 mg/L BAP, generating the maximum mean callus fresh weight (0.48 ± 0.10 g). The highest frequency of callus induction (88.00 ± 1.60 %) was obtained in solidified MS medium supplemented with 0.5 mg/L NAA + 2.0 mg/L BAP, producing the maximum mean fresh weight of callus (1.54 ± 0.27 g) and dry weight (0.90 ± 0.01 g). The results of the Phytochemical screenings of callus and dried leaf extracts indicated the presence of alkaloids, flavonoids, terpenoids, and saponins.

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