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

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.

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In vitro callus induction of gerbera from leaf explant

International Journal of Advanced Geosciences ◽

10.14419/ijag.v8i1.30000 ◽

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.

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Sterilization procedure and callus regeneration in black turmeric (Curcuma caesia)

Agricultural Science Digest - A Research Journal ◽

10.18805/ag.d-4714 ◽

2019 ◽

Vol 6 (49) ◽

Author(s):

A. S. Abubakar ◽

R. N. Pudake

Keyword(s):

Acetic Acid ◽

Callus Induction ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Surface Sterilization ◽

To Come ◽

Dichlorophenoxy Acetic Acid ◽

And Control ◽

Rhizome Explants

Sterilization procedure, media composition, explants selection and control of physical environment are critical for successful cultures and callus induction with surface sterilization being very challenging in most plants. Five different sterilization methods were evaluated to come up with the best for subsequent use to establish an in vitro regeneration method for the induction of callus in Curcuma caesia using excised leaf and rhizome explants. Murashige and Skoog (MS) media supplemented with various concentration of 2,4-Dichlorophenoxy acetic acid (2,4-D)/Indole-3-acetic acid (IAA) (0.5- 5.0mg/L), singly or in combination with Benzyl aminopurine (BAP)/Kinetin (KIN) (0.1-5.0mg/L), 0.3% sucrose and 0.08% agar were used. The result of the sterilization procedures showed 15% NaHClO3 (5min) + 70% Ethanol (30s) + 0.1% HgCl2 (5min) to be the most effective in controlling contamination in C. caesia among all the treatments tested. The response to callus induction was found to depend on the type of explants used and growth regulators combination. Leaf explants gave the highest percentage of callus induction. Highest percentage of callus induction (66.70%) was obtained in the growth regulator combination of 2, 4-D (0.5mg/L) + BAP (0.1mg/L) and least (14.29%) in IAA (2.0mg/L) + BAP (0.5mg/L). Equal and higher concentration of 2, 4-D + BAP of 5.0mg/L each also provided better result (40.00%). No callus was obtained in all the single concentration of 2, 4-D used.

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Micropropagation of Liriope muscari via leaf explant

The Journal of Agriculture of the University of Puerto Rico ◽

10.46429/jaupr.v83i3-4.3886 ◽

1969 ◽

Vol 83 (3-4) ◽

pp. 169-173

Author(s):

Keithley L. Amory ◽

John M. Gill

Keyword(s):

Acetic Acid ◽

Mass Production ◽

In Vitro Propagation ◽

Solid Medium ◽

Leaf Explants ◽

Ms Medium ◽

Isopentenyl Adenine ◽

Young Leaves ◽

Dichlorophenoxy Acetic Acid

Young leaves of Liriope muscari provide an ample source of explants for in vitro propagation in tropical countries where flowering is scarce. Leaves were induced to form calli on a solid medium containing Murashige and Skoog (MS) salts and vitamins, 3% sucrose, 0.7% agar, 1 mg/L 2,4-dichlorophenoxy- acetic acid (2, 4-D) and 1 mg/L 6-furfurylaminopurine (kinetin). Only the proximal segments of the leaves produced calli. These calli were induced to produce multiple plantlets on MS medium, 3% sucrose, 0.7% agar, and 10 mg/L N6 (2-isopentenyl) adenine (2 ip). It is possible to use leaf explants for in vitro mass production of Liriope. However, in variegated varieties, only green or white plants were produced, because of a chimera in the original tissue.

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Micropropagation, Callus Induction and Regeneration of Ginger (Zingiber officinale Rosc.)

Open Agriculture ◽

10.1515/opag-2020-0008 ◽

2020 ◽

Vol 5 (1) ◽

pp. 75-84

Author(s):

Seied Mehdi Miri

Keyword(s):

Acetic Acid ◽

Callus Induction ◽

Leaf Explants ◽

Multiple Shoots ◽

Zingiber Officinale ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Shoot Number ◽

Micro Propagation ◽

Dichlorophenoxy Acetic Acid

AbstractThe present study describes a protocol for micro-propagation, callus induction, and shoot regeneration of ginger (Zingiber officinale). The rhizomes were surface-sterilized with ethanol (70%) for 45 s, sodium hypochlorite (2.5%) for 10 min, and mercuric chloride (0.1%) for 10 min. Multiple shoots were induced from sprouting bud explants cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) combined with kinetin (Kin). The maximum shoot number was obtained from MS medium containing 10 mg/l BA with a mean of 20.6 shoots per explant. The leaf explants were cultured on MS medium supplemented with indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), Dicamba, or BA for callus culture. Green-red compact calli were induced using 2,4-D, Dicamba or BA. Also, BA successfully induced plant regeneration. The multiplied shoots that were transferred to the rooting medium (½MS supplemented with 0, 1 and 2 mg/l IAA, indole-3-butyric acid (IBA) or NAA) showed development of roots (100%). The rooted plantlets were transferred to pots containing a 1:1 mixture of cocopeat and perlite, and acclimatization was successful, resulting in 85% survival of the plantlets in the greenhouse.

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INFLUNCE OF GROWTH REGULATORS ON CALLUS INDUCTION OF CITRUS VOLKAMERIANA IN VITRO

IRAQI JOURNAL OF AGRICULTURAL SCIENCES ◽

10.36103/ijas.v47i3.561 ◽

2016 ◽

Vol 47 (3) ◽

Author(s):

Al- Khazali & Hamad

Keyword(s):

Acetic Acid ◽

Callus Induction ◽

Dry Weight ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Benzyl Adenine ◽

College Of Agriculture ◽

Citrus Volkameriana ◽

Dichlorophenoxy Acetic Acid

This research was conducted in the plant tissue culture Lab. College of Agriculture / University of Baghdad from February to October 2015. The aim of the study was investigating the influences of combinations of Naphthalene acetic acid (NAA) , Thidiazuron (TDZ) Spermidine (Spd. ) and 2,4-Dichlorophenoxy acetic acid (2,4-D) , Benzyl adenine (BA) on callus induction and adventitous shoot regeneration originated from cotyledon of Citrus volkameriana seeds. Seeds were disinfested with 0.1 % of HgCl2 for 15 minutes. The MS medium supplemented with (0.0,1.5 , 3.0 ) mg L-1 NAA in combination with (0.0, 0.05, 0.1) mg L-1 TDZ and (0.0, 0.5 ,1.0) mg L-1 Spd. and MS medium supplemented with (0.0, 1.5 , 3.0) mg L-1 2,4-D in combination with (0.0 ,1.0 , 2.0 ) mg L-1 BA and (0.0 ,0.5 , 1.0) mg L-1 Spd. the interaction between 1.5 mg L-1 NAA and (0.05 , 0.1) mg L-1 TDZ and the interaction between 3.0 mg L-1 NAA and (0.05 ,0.1) mg L-1 TDZ with all concentrations of Spd. gave the highest percentage of callus induction 100 % . While the MS medium supplemented with 3 mg L-1 of 2,4-D in combination with all concentrations of BA and spd. gave the highest percentage 100 % of callus induction. Results showed that MS medium supplemented with 1.5 mg L-1 NAA in combination with 0.1 mg L-1 TDZ and 1.0 mg L-1 spd. gave the highest values of fresh and dry weight of callus (668.8, 44.59 ) mg respectively . While the MS medium supplemented with 3 mg L-1 2,4-D in combination with 1.0 mg L-1 spd. And 0.0 mg L-1 BA gave the highest values of fresh and dry weight of callus (709.2 , 47.28 ) mg respectively.

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Callus Induction and Plant Regeneration in Rauvolfia Serpentina (L.) Benth Ex. Kurz.

Journal of Natural History Museum ◽

10.3126/jnhm.v24i1.2245 ◽

2009 ◽

Vol 24 ◽

pp. 82-88 ◽

Cited By ~ 1

Author(s):

Saraswoti Aryal ◽

Sanu Devi Joshi

Keyword(s):

Acetic Acid ◽

Callus Induction ◽

Coconut Milk ◽

Ms Medium ◽

Rauvolfia Serpentina ◽

History Museum ◽

Short Period ◽

Murashige Skoog ◽

Callus Tissue Culture

Rauvolfia serpentina (L.) ex. Kurz is an important medicinal plant. Callus induction and regeneration was studied from stem explant of in-vitro grown plant of Rauvolfia serpentina(L.) Benth. ex Kurz (Apocynaceae) on Murashige Skoog (1962) medium supplemented with 1mg/l 2,4-Dichlorophenocy acetic acid (2,4-D) and 1mg/l Kinetin (Kn). Vigorous growth of callus occurs after 4 weeks of culture. Callus was sub-cultured on Murashige and Skoog (MS) medium supplemented with different concentration of 2, 4-D (0.5-3.0 mg/l) and 10% coconut milk. Regeneration of plantlets occurred on MS medium containing 3 mg/1 of 2, 4-D and 10% coconut milk. These plantlets were rooted on MS medium supplemented with 1 mg/l IAA .The regenerated plantlets were able to grow on soil after short period ofacclimatization. Key words: Explant; In-vitro culture; MS medium; 2, 4 Dichlorophenoxy acetic acid; Kinetin; Callus; Tissue culture; Coconut milk. Journal of Natural History Museum Vol. 24, 2009 Page: 82-88

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Influence of growth regulators and explants on shoot regeneration in carnation

Horticultural Science ◽

10.17221/1/2009-hortsci ◽

2009 ◽

Vol 36 (No. 4) ◽

pp. 140-146 ◽

Cited By ~ 4

Author(s):

J.K. Kanwar ◽

S. Kumar

Keyword(s):

Acetic Acid ◽

Shoot Regeneration ◽

Growth Regulators ◽

Dianthus Caryophyllus ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Shoot Bud ◽

Dichlorophenoxy Acetic Acid ◽

Number Of Shoots

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.

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

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha4028248 ◽

2012 ◽

Vol 40 (2) ◽

pp. 140 ◽

Cited By ~ 2

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.

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

Kongunadu Research Journal ◽

10.26524/krj202 ◽

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.

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In Vitro Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey

10.5772/intechopen.94378 ◽

2020 ◽

Author(s):

Nurşen Çördük ◽

Cüneyt Aki

Keyword(s):

Acetic Acid ◽

Medicinal Plant ◽

In Vitro Propagation ◽

Leaf Explants ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Northwestern Turkey ◽

Helen Of Troy ◽

Endemic Medicinal Plant

Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regeneration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber.

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