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 Adaptation of in vitro regeneration protocol for Brazilian wheat genotypes

2018 ◽  
Vol 48 (11) ◽  
Author(s):  
Eduardo André Roesler ◽  
Ernandes Manfroi ◽  
Andréa Morás ◽  
Dielli Aparecida Didoné ◽  
Magali Ferrari Grando ◽  
...  
Keyword(s):  
Callus Induction ◽  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Regeneration Capacity ◽  
Target Region ◽  
Wheat Genotypes ◽  
Regeneration Efficiency ◽  
Callus Induction Medium

ABSTRACT: The availability of an efficient protocol for in vitro regeneration is imperative for genetic transformation. Using genotypes adapted to the target region as a transgenic platform accelerates the development of cultivars. Therefore, this study aimed to adapt an in vitro regeneration protocol for Brazilian wheat genotypes. For this purpose, the in vitro regeneration capacity of immature embryos from six Brazilian wheat genotypes using two protocols of regeneration of somatic embryos was analysed. Furthermore, combinations of 2,4-D and picloram in the callus induction medium were tested in order to improve regeneration efficiency. Genotypes with higher regeneration efficiency were BR18-Terena and PF020037, yielding 0.42 and 1.13 shoots per explant using the Hu and the Wu protocol, respectively. Adding 1mgL-1 2,4-D in the callus induction medium was the most favourable, producing 3.73 and 3.07 shoots per explant for PF020037 and BR18-Terena, respectively. In conclusion, a protocol for regeneration for two Brazilian wheat genotypes recommended and developed to be cultivated at the Cerrado region has been adapted.

scholarly journals Induction of Morphogenic Callus and In Vitro Regeneration of Sugarcane (Saccharum Officinarum L.)

2021 ◽  
Vol 5 (1) ◽  
pp. 61-67
Author(s):  
Sitti Inderiati ◽  
FNU Yanti ◽  
Eka Ria Mentari
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
In Vitro Regeneration ◽  
Shoot Proliferation ◽  
Saccharum Officinarum ◽  
Induction Medium ◽  
Ms Medium ◽  
Shoot Initiation ◽  
Morphogenic Callus

In vitro propagation is a method to produce massive healthy new planting materials quickly. An experiment was carried out for morphogenic callus induction and regeneration of a domestic sugarcane variety. The Explants used was an inert folded leaf and incubated on modified MS medium augmented with 1 mg/l, 2.5 mg/l, and 5 mg/l of 2,4-D for callus induction. The leaf calluses were subcultured on MS medium enriched with different growth regulators for shoot initiation and multiplication. The highest percentage of callus formation was achieved in the medium containing 2.5 mg/l of 2,4-D, while the fastest callus initiation was noticed in MS medium supplemented with 5 mg/l 2,4-D, and maximum proliferation and the morphogenic response of callus were obtained in 3rd subculture. Two types of callus observed on the induction medium were dry nodular friable and smooth compact. This highly morphogenic callus was white to white creamy in color and easy to separate.   The highest shoot proliferation rate was found on the medium containing 2 mg/l Kinetin + 1 mg/l IAA and no growth was noticed on the medium containing Kinetin alone. Therefore, the study suggests that the growth hormone of cytokinin in combination with auxin is necessary for in vitro regeneration of sugarcane callus culture.

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

2009 ◽  
Vol 24 ◽  
pp. 82-88 ◽  
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

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 In vitro bulblet regeneration from immature embryos of endangered and endemic Muscari azureum

2011 ◽  
Vol 63 (1) ◽  
pp. 209-215 ◽  
Author(s):  
S. Uranbey
Keyword(s):  
Callus Induction ◽  
High Frequency ◽  
Ornamental Plant ◽  
Immature Embryos ◽  
Induction Medium ◽  
Ms Medium ◽  
Culture Initiation ◽  
Mineral Salts ◽  
Font Color

A high frequency of bulblet regeneration was achieved for the endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on a callus induction medium consisting of N6 mineral salts and vitamins, 400 gL-1 casein + 40 gL-1 sucrose + 2 mgL-1 L-proline, 2 mgL-1 2,4-D and 2 gL-1 Gelrite. Then the embryogenic callus clusters were transferred to a bulblet induction medium consisting of MS mineral salts and vitamins containing different concentrations and combinations of BAP, KIN, TDZ, Zeatin, IAA, NAA, 30 gL-1 sucrose and 7 gL-1 agar. Prolific bulblet multiplication (over 13 bulblets/embryo) was achieved from immature embryos after 5-6 months of culture initiation. Well-developed bulblets were excised and individually rooted on ? strength MS medium supplemented with 1 mgL-1 IBA, 0.5 gL-1activated charcoal, 20 gL-1sucrose and 6 gL-1agar and acclimatized. <br><br><font color="red"><b> This article has been retracted. Link to the retraction <u><a href="http://dx.doi.org/10.2298/ABS150608072E">10.2298/ABS150608072E</a><u></b></font>

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

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.

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 Mature Embryo-Based in vitro Regeneration of Indica Rice Cultivars for High Frequency Plantlets Production

2017 ◽  
Vol 20 (2) ◽  
pp. 81-87
Author(s):  
HN Barman ◽  
ME Hoque ◽  
RK Roy ◽  
PL Biswas ◽  
MAI Khan ◽  
...  
Keyword(s):  
Callus Induction ◽  
High Frequency ◽  
Indica Rice ◽  
In Vitro Regeneration ◽  
Mature Embryo ◽  
Ms Medium ◽  
Rice Varieties ◽  
Growing Medium ◽  
Regeneration Efficiency

The study was conducted at Biotechnology Division of Bangladesh Rice Research Institute (BRRI) to investigate the effects of plant growing medium and plant growth regulator (PGR) for the callus induction and high frequency plantlets regeneration of indica rice. Ten indica rice varieties viz. BR5, BR11, BRRI dhan28, BRRI dhan29, BRRI dhan33, BRRI dhan41, BRRI dhan47, BRRI dhan48, BRRI dhan49 and BRRI dhan50 were cultured on MS, N6 and LS media. The MS medium was found better for callus induction as compared to N6 and LS media. Among the tested varieties BRRI dhan48 induced the highest percent and best quality callus. Interaction effects of BRRI dhan48 to MS medium yielded 71.55% callus induction. The regeneration efficiency of BRRI dhan48 was tested on MS medium supplemented with different combination of NAA plus BAP and NAA plus kinetin. MS medium supplemented with 2.0 mg L-1 NAA and 2.0 mg L-1 Kn was found the best in respect of percent regenerated (76.67%) plantlet as well as for the growth of plantlets in vitro.Bangladesh Rice j. 2016, 20(2): 81-87

The Effects of Cytokinins on Plant Regeneration of Vetiver Grass (Vetiveria nemoralis A. Camus cv. Roiet)

2015 ◽  
Vol 804 ◽  
pp. 259-262
Author(s):  
Chonnikarn Khunchuay ◽  
Kanokporn Sompornpailin
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Shoot Number ◽  
Induction Experiment

The optimum ratios of auxin and cytokinin are necessary for callus induction and plant regeneration. This ratio is a key function involving in the promoting cell division and proliferation in tissue culture. The axillary buds of in vitro plantlets fromVetiveria nemoralisA. Camuscv. Roiet were used as explants for the callus induction experiment. These explants were cultured on Murashige & Skoog (MS) medium [1] supplemented with various combinations of auxins and cytokinins. Under this experimental study, the highest frequency of callus induction was found on MS medium supplemented with 2 mgL-1α-naphthalene acetic acid (NAA) and 1 mgL-12-furanylmethyl-1H-purine-6-amine (kinetin) (62.5%). On the other hand the combination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 6-benzylaminopurine (BAP) was toxicity to this explants. All culturing explants were dead and no calli appearance. The calli derived from each medium were transferred into the same regeneration medium (MS with 1 mgL-1NAA and 2 mgL-1BAP). After culturing on regeneration medium, calli induced from the highest callus induction medium have shown high frequencies of regeneration and also shoot number per callus (58.33% and 7.1 shoots).

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