scholarly journals Improving Adventitious Shoot Regeneration and Transient Agrobacterium-Mediated Transformation of Apricot (Prunus armeniaca L.) Hypocotyl Sections

Agronomy ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1338
Author(s):  
Cristian Pérez-Caselles ◽  
Lydia Faize ◽  
Lorenzo Burgos ◽  
Nuria Alburquerque
Keyword(s):  
Adventitious Shoot ◽  
Prunus Armeniaca ◽  
Vacuum Infiltration ◽  
Dichlorophenoxyacetic Acid ◽  
Transient Transformation ◽  
Regeneration Medium ◽  
Transformation Protocol ◽  
Efficient Regeneration ◽  
Prunus Armeniaca L ◽  
2,4 Dichlorophenoxyacetic Acid

The improvement of previously described protocols for the regeneration of shoots from ‘Canino’ mature seed hypocotyl slices has been accomplished. The effects of different factors such as the part of the hypocotyl used, vacuum-infiltration, 2,4-Dichlorophenoxyacetic acid pulse, vacuum-infiltration and sonication on regeneration and transient transformation were analyzed. When the three slices obtained from the hypocotyls were evaluated separately on regeneration medium, the highest percentages of regenerating explants were achieved in the part close to the epicotyl and in the central part. On the other hand, sonication of the explants for 30 s followed by vacuum-infiltration during Agrobacterium infection for 20 min allowed for an increase in the transformation events. The application of these modifications to the procedure increased the regeneration efficiencies, and transient transformation events and may reduce the frequency of failed experiments. An efficient regeneration/transformation protocol could facilitate its use as a biotechnological technique for apricot breeding.

scholarly journals CALLUS INDUCTION ON BANANA FLOWER’S EXPLANT IN VITRO USING 2,4-DICHLOROPHENOXYACETIC (2,4-D)

2018 ◽  
Vol 22 (2) ◽  
pp. 66
Author(s):  
RINDANG - DWIYANI ◽  
HESTIN - YUSWATI ◽  
UTAMI -
Keyword(s):  
Acetic Acid ◽  
Callus Induction ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Regeneration Medium ◽  
Indirect Organogenesis ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Banana Cultivar

ABSTRACT  The objective of the study was to obtain the best 2,4-D concentration on callus induction of the banana flowers in banana propagation using indirect organogenesis method. Kesuna, local banana cultivar obtained from Sembung Gede, Tabanan was used as explant material. Callus induction was performed using 2,4-Dichlorophenoxyacetic acid with concentration of 0; 0.5; 1.0; 1.5 and 2.0 ppm. Each treatment was represented by 3 bottles and each bottle was planted with 3 explants, so each treatment was represented by 9 explants of banana flowers. The results showed that the concentration of 2.0 ppm 2.4-D induced callus with the fastest time and gave the highest percentage of the explants producing callus. The calluses were subsequently subcultured into regeneration medium using 0.5 mg/L Benzylaminopurine (BAP) and 0.005 mg/L Napthaleneaceticacid (NAA). The calluses were subsequently sub-cultured into a regeneration medium using 0.5 ppm (BAP) and 0.005 ppm Naphthalene acetic acid (NAA) to induce shoots and roots and performed plantlets.   Keywords: 2,4-Dichlorophenoxyacetic acid, banana’s flowers, callus

scholarly journals Regeneration of Ornamental Cherry (Prunus) Taxa from Mature Stored Seed

HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 745-748 ◽  
Author(s):  
Karen E. Hokanson ◽  
Margaret R. Pooler
Keyword(s):  
Callus Formation ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Adventitious Shoot Regeneration ◽  
Potential System ◽  
Regeneration In Vitro ◽  
2,4 Dichlorophenoxyacetic Acid

Callus formation and adventitious shoot regeneration in vitro from mature stored seed were evaluated in eight ornamental cherry (Prunus) taxa: P. campanulata Maxim., P. maackii Rupr., P. sargentii Rehd., P. serrula Franch., P. serrulata Lindl., P. subhirtella Miq., P. virginiana L., and P. yedoensis Matsum. Several portions of the embryo (cotyledons and hypocotyl sections) and nine combinations of growth regulators (BA, 2,4-D, IBA, NAA, and TDZ) were compared. Effects of embryo portions and growth regulator treatments were generally small within taxa, but shoot formation differed among taxa. About 20% to 50% of the embryos from P. virginiana and P. serrula and ≈5% to 30% of those from P. maackii produced shoots. The other taxa generally did not produce shoots. Regeneration from mature stored seed in the responsive taxa represents a potential system for genetic transformation. Chemical names used: 6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); thidiazuron (TDZ).

scholarly journals 099 Indirect and Direct Regeneration of Kalmia latifolia

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 458D-458
Author(s):  
Mark H. Brand
Keyword(s):  
Shoot Regeneration ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Direct Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Kalmia Latifolia ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Optimal Medium ◽  
Silver Dollar

To introduce desirable trait genes into Kalmia latifolia, efficient adventitious shoot regeneration methods are needed. Silver Dollar (S$) callus induction and growth in the dark was compared on Woody Plant (WP) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (1, 5, 10, 20 μM) or naphthaleneacetic acid (NAA) (1, 10, 20, 40 μM) with and without 5 μM isopentenyladenine (2iP). Both 2,4-D and NAA produced >450 mg of callus from leaf explants in 8 weeks. The addition of 2iP tripled growth for 2,4-D and doubled growth for NAA. Greatest callus growth was obtained on 20-40 μM NAA or 5-20 μM 2,4-D. Shoot regeneration on callus was achieved on WP medium containing 30 μM 2iP or 1 μM thidiazuron (TDZ), but a combination of the two was best, with 68% of dark-grown calli regenerating shoots in 4 weeks. 26% more dark-grown calli regenerated shoots than light-grown calli. The type of auxin (2,4-D or NAA) used to grow the calli did not affect shoot regeneration. For direct shoot regeneration, S$ leaf explants were tested on WP medium containing 5, 15, 30, 45 and 60 μM 2iP. The addition of 1 μM indole-3-butyric acid (IBA) doubled the percentage of leaves that regenerated shoots. 2iP concentrations between 15 and 45 μM supported excellent shoot regeneration, but optimal regeneration (95% of explants, 5.1 shoots/leaf) occurred on 30 μM 2iP+1 μM IBA. Leaf explants of six cultivars were grown on optimal medium with shoot regeneration ranging from 17% to 93% of leaves and 1.8 to 8.2 shoots per leaf, depending on the cultivar.

scholarly journals PCIB Enhances Regeneration of Ipomoea cordatotriloba Dennstedt Leaf Explants and Protoplasts

HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 327-328
Author(s):  
Ruth S. Kobayashi ◽  
John C. Bouwkamp ◽  
Stephen L. Sinden
Keyword(s):  
Abscisic Acid ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Regeneration Medium ◽  
Ipomoea Cordatotriloba ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Chlorophenoxyisobutyric Acid

Leaf callus of Ipomoea cordatotriloba was initiated by culturing explants on Linsmaier and Skoog medium containing 3 g yeast extract/liter, 18.9 μm ABA, 2.3 μm 2,4-D, and 0.15 m sucrose. Calluses were transferred to Murashige and Skoog media containing 17.8 μm BA and 0, 1, 10, or 100 μm PCIB. The number of shoots from calluses grown on medium containing 10 μm PCIB increased significantly, and the percentage of calluses exhibiting shoot regeneration almost doubled compared to calluses grown on regeneration medium without PCIB. Protoplasts isolated from stem and petiole tissues of in vitro-grown plants were cultured in Kao and Michayluk 8p medium to the callus stage. Calluses (4-6 mm) were transferred to the callus induction and regeneration media used to regenerate leaf-explant callus. Of the protoplast-derived calluses cultured on media containing 10 or 100 μm PCIB, ≈13% and 18%, respectively, regenerated shoots after 2 months; none regenerated on the medium without PCIB. Chemical names used: abscisic acid (ABA); 2,4-dichlorophenoxyacetic acid (2,4-D); N6-benzyladenine (BA); α -p-chlorophenoxyisobutyric acid (PCIB).

scholarly journals Anatomical study of Racosperma (ex-Acacia) mangium tissues cultured in vitro

2000 ◽  
Vol 43 (1) ◽  
Author(s):  
N. K. Takemori ◽  
R. Marschner ◽  
M. Quoirin ◽  
C. Bona ◽  
F. Zanette
Keyword(s):  
Acacia Mangium ◽  
Anatomical Study ◽  
Adventitious Shoot ◽  
Basic Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Adventitious Shoot Regeneration ◽  
Meristematic Cells ◽  
Parenchyma Cells ◽  
2,4 Dichlorophenoxyacetic Acid

In order to achieve adventitious shoot regeneration from young tissues of Racosperma mangium, several combinations of growth regulators were tested. Kinetin (KIN) and a -naphthalenacetic acid (NAA), thidiazuron (TDZ) and NAA or indole-3-butyric acid (IBA), 4-CPPU and 2,4-dichlorophenoxyacetic acid (2,4-D) were added to basic medium composed of MS salts, MS vitamins and 3% sucrose. Sections of hypocotyls and cotyledons of young seedlings were cultivated on these media. Calli developed on different media and, after two to three months, some of them regenerated organized structures. Histological studies revealed hypertrophy and hyperplasy of medular and cortical parenchyma cells of hypocotyl and cotyledon parenchyma. Formation of meristematic cells and traqueary elements from parenchyma was observed during in vitro culture. In some cases, nodules of traqueary elements were formed. The highest differentiation into meristematic cells was found in cotyledon tissues cultured on 4-CPPU (8.07 µM) and 2,4-D (1.13 µM).

scholarly journals Simplified Regeneration Protocol for Cycas revoluta Thunb. Mature Zygotic Embryos

2015 ◽  
Vol 7 (1) ◽  
pp. 62-65
Author(s):  
Rohangiz NADERI ◽  
Khadije MOHAISENI ◽  
Jaime A. TEIXEIRA DA SILVA ◽  
Mansour OMIDI ◽  
Behjat NADERI
Keyword(s):  
Zygotic Embryo ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Indirect Organogenesis ◽  
Rooted Plantlet ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Cycas Revoluta

Mature zygotic embryos of Cycas revoluta Thunb. were used as explants to investigate direct and indirect organogenesis. Explants were incubated on half-strength Murashige and Skoog (½ MS) basal medium supplemented with various plant growth regulators, singly or in combination (all at 0.5 mg l-1): 6-benzyladenine (BA), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), Kin×2,4-D, BA×Kin and BA×2,4-D. Cultures were placed at a low light intensity (4 µmol m-2 s-1 PPFD). Adventitious shoot regeneration was observed in the presence of 0.5 mg l-1BA after 35 days. The highest number of direct and indirect shoots per zygotic embryo was 3.67 and 29.67, respectively. Roots were induced on indirect shoots by continuous culture on rooting medium (½ MS,‏ 0.1 mg l-1 1-naphthaleneacetic acid) and hardened successfully in perlite. Each rooted plantlet with pinnate leaves and a primary tap root was individually isolated and acclimatized 185 days after the beginning of culture, with a 10% success rate.

scholarly journals Shoot Regeneration in Vitro from Diverse Genotypes of Sweetpotato and Multiple Shoot Production per Explant

HortScience ◽  
1995 ◽  
Vol 30 (5) ◽  
pp. 1074-1077 ◽  
Author(s):  
Ramana M. Gosukonda ◽  
C.S. Prakash ◽  
Ananta Porobo Dessai
Keyword(s):  
Shoot Regeneration ◽  
Ipomoea Batatas ◽  
Multiple Shoots ◽  
Adventitious Shoot ◽  
Multiple Shoot ◽  
Dichlorophenoxyacetic Acid ◽  
Horizontal Orientation ◽  
Shoot Production ◽  
2,4 Dichlorophenoxyacetic Acid

Studies were conducted to improve adventitious shoot regeneration in sweetpotato [Ipomoea batatas (L.) Lam.], specifically to extend the protocol to many genotypes and to elicit production of multiple shoots per explant. The use of a two-stage procedure where excised petioles were incubated on Murashige and Skoog (MS) (1962) medium with 2,4-D (0.2 mg·liter–1) for 3 days and transferred to a second medium containing MS salts with thidiazuron and 2iP (0.05 mg·liter–1 each) resulted in shoot regeneration from eight of 13 genotypes tested, including elite sweetpotato cultivars such as `Jewel' and `Rojoblanco'. PI 318846-3 was the most regenerable genotype, with up to 77% of explants producing one to three shoots per explant. The orientation of the petiole on the nutrient medium was critical; those placed vertically inverted developed multiple shoots. Wounding explants through epidermal peeling with normal horizontal orientation of the explants during incubation also resulted in multiple shoot production (about three shoots per explant). Interference with auxin transport due to explant inversion or wounding may have stimulated increased shoot induction. Chemical names used: 2,4 dichlorophenoxyacetic acid (2,4-D); N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (thidiazuron); N6-(2-isopentenyl) adenine (2iP).

In Vitro Shoot Regeneration from Callus Derived from Marubakaido Apple Rootstock under Different Aluminium Concentrations

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva
Keyword(s):  
Shoot Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Apple Rootstock ◽  
Under Five ◽  
Significant Difference ◽  
Previously Treated ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.

Characterization of the Reproductive Mode in Guayule In Vitro

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig
Keyword(s):  
Field Experiments ◽  
Reproductive Mode ◽  
Dichlorophenoxyacetic Acid ◽  
Initial Field ◽  
Breeding Lines ◽  
Reproductive Modes ◽  
Desert Southwest ◽  
2,4 Dichlorophenoxyacetic Acid

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.

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