scholarly journals The effect of auxins in inducing organogenesis or somatic embryogenesis in mature sunflower zygotic embryo derived apex

2020 ◽  
Vol 48 (1) ◽  
pp. 150-161
Author(s):  
Adriana AURORI ◽  
Imola MOLNAR ◽  
Elena RAKOSY-TICAN
Keyword(s):  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Mature Stage ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Axillary Shoots ◽  
Mature Embryos ◽  
Cytokinin Treatment ◽  
2,4 Dichlorophenoxyacetic Acid

Induction of shoots or of somatic embryos is the key step for gaining the morphogenetic potential in sunflower (Helianthus annuus L.), species known as recalcitrant to in vitro regeneration. In the immature zygotic embryo derived tissues or in other juvenile tissues resulted from seedlings, the acquisition of the competence for regeneration can be achieved directly by cytokinin treatment or by preconditioning the explants on cytokinin containing medium. In this paper is presented a new type of explant for sunflower in vitro culture, consisting of the apex with primordial leaves, resulted from ungerminated mature zygotic embryo, in which a specific morphogenetic response was triggered by the exogenously applied auxins. Among the auxins tested, indole-3-acetic acid, indole-3-butyric acid and 1-naphthaleneacetic acid are inducers of an organogenetic response, apical/axillary shoots and adventitious buds being regenerated while 2,4-dichlorophenoxyacetic acid, 3,6-dichloro-2-methoxybenzoic acid and 4-amino-3,5,6-trichloropicolinic acid led to somatic embryo formation. Among the auxins tested only 4-amino-3,5,6-trichloropicolinic acid sustains the embryos development up to mature stage. A high amount of sucrose (120 g L-1) supplied during the auxin treatment promotes the maturation of the embryos directly on the induction medium for all tested auxins with embryogenic effect. These findings show that regardless of the type of morphogenetic response aimed in sunflower meristematic tissues resulted from mature embryos, the presence of auxins is mandatory.

scholarly journals Shoot Proliferation, Callus Induction And Plant Regeneration In Tripsacum Laxum Nash

2021 ◽  
Author(s):  
Yuping Xiong ◽  
Jinhui Pang ◽  
Kunlin Wu ◽  
Jaime A. Teixeira Silva ◽  
Xinhua Zhang ◽  
...  
Keyword(s):  
Peat Soil ◽  
Shoot Proliferation ◽  
Multiple Shoots ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Rooting Medium ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract The peduncles of Tripsacum laxum Nash were used as explants to induce axillary shoots. Multiple shoots were proliferated on Murashige and Skoog (MS) medium to establish, for the first time, efficient shoot proliferation and plant in vitro regeneration systems. Optimal shoot proliferation medium was MS with 3.0 mg/L 6-benzyladenine (BA) and 0.2 mg/L α-naphthaleneacetic acid (NAA), resulting in a shoot proliferation coefficient of 11.0 within 45 d. Optimal rooting medium was MS with 0.1 mg/L NAA and/or 0.1 mg/L indole-3-butyric acid (IBA), inducing 100% root formation from shoots within 30 d. When young roots, leaf sheaths and shoot bases were used as explants, MS medium with 1.0 mg/L thidiazuron (TDZ) and 0.2 mg/L BA induced most shoots, with the least callus. Shoot bases induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), while leaf sheaths induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L BA. Rooted plantlets showed 99.3% survival when transplanted into a substrate of vermiculite: peat soil (1:3, v/v).

scholarly journals Application of Artificial Neural Network for Modeling and Studying In Vitro Genotype-Independent Shoot Regeneration in Wheat

2020 ◽  
Vol 10 (15) ◽  
pp. 5370 ◽  
Author(s):  
Mohsen Hesami ◽  
Jorge A. Condori-Apfata ◽  
Maria Valderrama Valencia ◽  
Mohsen Mohammadi
Keyword(s):  
Neural Network ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Generalized Regression Neural Network ◽  
Naphthaleneacetic Acid ◽  
Regeneration Frequency ◽  
Artificial Neural ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

Optimizing in vitro shoot regeneration conditions in wheat is one of the important steps in successful micropropagation and gene transformation. Various factors such as genotypes, explants, and phytohormones affect in vitro regeneration of wheat, hindering the ability to tailor genotype-independent protocols. Novel computational approaches such as artificial neural networks (ANNs) can facilitate modeling and predicting outcomes of tissue culture experiments and thereby reduce large experimental treatments and combinations. In this study, generalized regression neural network (GRNN) were used to model and forecast in vitro shoot regeneration outcomes of wheat on the basis of 10 factors including genotypes, explants, and different concentrations of 6-benzylaminopurine (BAP), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), zeatin, and CuSO4. In addition, GRNN was linked to a genetic algorithm (GA) to identify an optimized solution for maximum shoot regeneration. Results indicated that GRNN could accurately predict the shoot regeneration frequency in the validation set with a coefficient determination of 0.78. Sensitivity analysis demonstrated that shoot regeneration frequency was more sensitive to variables in the order of 2,4-D > explant > genotype < zeatin < NAA. Results of this study suggest that GRNN-GA can be used as a tool, besides experimental approaches, to develop and optimize in vitro genotype-independent regeneration protocols.

scholarly journals An efficient in vitro regeneration system via callus from hypocotyl and root of Iris laevigata

2020 ◽  
Author(s):  
Fengyi Li ◽  
Lijuan Fan ◽  
Haijing Fu ◽  
Yujia Liu ◽  
Ling Wang
Keyword(s):  
In Vitro Regeneration ◽  
Genetic Material ◽  
Reproductive Capacity ◽  
Penicillin G ◽  
Induction Medium ◽  
Multiplication Rate ◽  
Regeneration System ◽  
Root Explants ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract Background Iris laevigata is an ornamental plant with strong cold resistance. However, its low reproductive capacity limits its landscape applications. The I. laevigata wild genetic resources also need to be protected. In order to develop an effective regeneration system, the optimum agar concentration for the induction medium was determined. Two explants (hypocotyl and root) were then cultured on medium containing different concentrations of plant growth regulator (PGR). In addition, three antibiotics were evaluated for controlling endophyte contamination. Results The highest induction rate (75.00%) was obtained from hypocotyl explants on Murashige and Skoog salt mixture (MS) medium containing 6-benzylaminopurine (6-BA) 0.5 mg L-1 + 2,4-Dichlorophenoxyacetic acid (2,4-D) 1.0 mg L-1 + 1-naphthylacetic acid (NAA) 0.4 mg L-1. The medium containing 6-BA 0.5 mg L-1 + 2,4-D 0.5 mg L-1 + NAA 0.2 mg L-1 achieved the greatest multiplication rate (73.33%). Media containing indole-3-butyric acid (IBA) 0.5 mg L-1 + 6-BA 1.5 mg L-1 + NAA 1.0 mg L-1 achieved the highest differentiation rate (39.72%) for hypocotyl induced calli. Medium containing 6-BA 2.0 mg L-1 + NAA 0.4 mg L-1 + kinetin (KT) 1.0 mg L-1 resulted in the highest differentiation rate (49.52%) for root induced calli. One hundred mg L-1 penicillin G resulted in the optimal rate for reducing endophyte contamination. Conclusions The research determined the optimum PGR concentrations for inducting and multiplying I. laevigata calli from hypocotyl and root explants and a satisfactory means for control endophyte contamination. This research will result in the efficient and reliable reproduction of I. laevigata for landscape applications, genetic development of new Iris varieties, and preservation of the wild genetic material.

scholarly journals Response of Different Explants for Callus Induction in Cucumber

2021 ◽  
Vol 2 (5) ◽  
pp. 71-75
Author(s):  
Hasina Sultana ◽  
Lutfun Nahar ◽  
M. Mofazzal Hossain ◽  
Totan Kumar Ghosh ◽  
Md. Sanaullah Biswas
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Disc ◽  
Dry Weight ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Regenerate Shoot ◽  
2,4 Dichlorophenoxyacetic Acid

In vitro regeneration of cucumber is relatively difficult for genetic improvement. In this regard, different concentrations of growth regulators and three types of explants (cotyledon, hypocotyl and leaf disc) were investigated for their efficiency on callus induction potential. Among different explants explored for callus induction with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), leaf disc responded earlier (4.67 days) and showed higher percentage of callus induction (91.50%) with 2 mg/l 2,4-D supplemented Murashige and Skoog (MS) media. The same concentration of 2,4-D resulted in the maximum callus fresh (0.56 g) and dry weight (0.39 g) from leaf disc explant. Then the callus was transferred to untreated, 2.0 mg/l BAP + 0.2 mg/l NAA + 1.0 mg/l Kn, 2.0 mg/l BAP + 1.0 mg/l NAA + 1.0 mg/l Kn and 2.0 mg/l BAP + 1.5 mg/l NAA + 1.0 mg/l Kn fortified MS medium. After transferring the callus of different explants to shoot regeneration media containing different concentrations of 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA) and Kinetin (Kn), only cotyledon callus started to regenerate shoot. The combination of BAP (2 mg/l) + NAA (0.2 mg/l) + Kn (1 mg/l) showed highest shoot regeneration percentage (67.77%) and the maximum number of shoots (5.12) per explant were recorded in the treatment combination of 2 mg/l BAP + 0.2 mg/l NAA + 1 mg/l Kn. These results provided a basis for the optimization of the callus induction protocol of cucumber for genetic transformation.

scholarly journals Optimization of Callogenesis/Caulogenesis Induction Protocol in Saffron Plant (Crocus sativus L.) Using Response Surface Methodology

2021 ◽  
Vol 12 (4) ◽  
pp. 4731-4746
Keyword(s):  
Callus Formation ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Crocus Sativus ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Indole Butyric Acid ◽  
2,4 Dichlorophenoxyacetic Acid

The Crocus sativus, an endangered medicinal and aromatic plant in Morocco, has a low propagation rate in natural conditions and, therefore, an efficient method for in vitro propagation is required. This study investigated the effects of various hormones on the induction of callogenesis and callogenesis in C. sativus corms using the Box-Behnken experimental design. The best shoot formation was obtained with Murashige and Skoog fortified with 3 mg/L 6-Benzylaminopurine. On the other hand, callus formation was obtained with 3 mg/L 1-Naphthaleneacetic Acid or 3 mg/L 2,4-Dichlorophenoxyacetic Acid. However, a combination of 3 mg/L 6-Benzylaminopurine, 1.056 mg/L Indole Butyric Acid, and 3 mg/L 2,4-Dichlorophenoxyacetic Acid allows 50% caulogenesis and 60% callogenesis. The in vitro regeneration system could be utilized for both conservation and largescale multiplication of Crocus sativus corms.

Somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of allium altaicum pall.

2018 ◽  
Vol 22 (03) ◽  
pp. 82-88
Author(s):  
Zavzandulam М ◽  
Buyanchimeg B ◽  
Enkhchimeg V
Keyword(s):  
Callus Induction ◽  
Zygotic Embryo ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Red Data Book ◽  
Data Book ◽  
2,4 Dichlorophenoxyacetic Acid

Altain onion (Allium altaicum Pall.) grows wildly under different ecological conditions and one of the listed rare plant in Red Data Book of Mongolia. Allium altaicum pall belong to a member of the onion family (Alliaceae) and has been used for both culinary and traditional medicine and a perennial herb.The purpose of this research is to get micropropogated plants in in vitro condition from Mongolian the Allium altaicum Pall tissue culture. Allium altaicum Pall. regeneration from zygotic embryo was 70% in MS medium with 0.5 mg/l 1-Naphthaleneacetic acid, 0.2 mg/l kinetin compare to control. Convenient condition for primary callus induction observed in MS medium with 1 mg/l 2,4-dichlorophenoxyacetic acid, 0.6 mg/l 6-benzylaminopurine, 2mg/l glycine by 50.4%. Regeneration of callus induction was 61.3% and somatic embryos formed plantlets on regeneration 0.1 мг/л 2,4-D 0.1 mg/l 2,4-dichlorophenoxyacetic acid, 1 мг/л BAP 1 mg/l 6-benzylaminopurine.

Somatic embryogenesis in American chestnut

1991 ◽  
Vol 21 (11) ◽  
pp. 1698-1701 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko ◽  
B. A. Watson-Pauley
Keyword(s):  
Somatic Embryos ◽  
Zygotic Embryo ◽  
American Chestnut ◽  
Induction Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Free Medium ◽  
Embryo Cultures ◽  
Proembryogenic Masses ◽  
2,4 Dichlorophenoxyacetic Acid

Cultures were initiated from developing ovules and excised embryos of American chestnut (Castaneadentata (Marsh.) Borkh.), collected from five source trees on three dates during early and middle stages of fruit development. Explants were cultured initially on semisolid induction medium containing 0.25 mg/L benzyladenine and either 6 mg/L naphthaleneacetic acid or 4 mg/L 2,4-dichlorophenoxyacetic acid for 1 or 2 weeks. Then they were either transferred to hormone-free medium or medium with 0.25 mg/L benzyladenine or maintained on the original induction media. Ovules collected from three of the five trees 6 or 7 weeks postanthesis produced embryogenic cultures. Those pulsed for 1 or 2 weeks on auxin-containing medium prior to transfer to media without auxin produced multiple somatic embryos directly from the radicle end of the zygotic embryo. Cultures maintained on auxin-supplemented media initially produced proembryogenic masses, which formed globular and heart-stage embryos as they aged. Transfer of clusters of somatic embryos from auxin-supplemented media to hormone-free medium promoted maturation of embryos to the cotyledon stage.

Intensification de la régénération du pois (Pisum sativum L.), par le thidiazuron, via la formation de structures caulinaires organogènes

1997 ◽  
Vol 75 (3) ◽  
pp. 492-500 ◽  
Author(s):  
Delphine Popiers ◽  
Frédéric Flandre ◽  
Brigitte S. Sangwan-Norreel
Keyword(s):  
Pisum Sativum ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Cotyledonary Nodes ◽  
Pisum Sativum L ◽  
Embryo Axes ◽  
Initial Medium ◽  
Second Wave ◽  
Mature Seeds

In vitro regeneration of pea (Pisum sativum L.), a regeneration recalcitrant legume, was optimised using thidiazuron. Buds were initiated from the meristems of the cotyledonary nodes of embryo axes, isolated from mature seeds, and subcultured on Murashige and Skoog medium supplemented with 13.3 μM 6-benzylaminopurine, 16.1 μM α-naphthaleneacetic acid, and 0.2 μM 2,3,5-triiodobenzoic acid. Proliferation of buds was preceded by the formation of white nodular-like protrusions. These structures were cut transversally in fine slices and subcultured on the same medium or in presence of thidiazuron that produces a second wave of secondary budding. The best results (90–110 buds per expiant) were obtained with 10 μM thidiazuron. The capacity of regeneration was genotype independent and reproducible. Buds elongated on the initial medium, then formed roots in presence of 5.37 μM α-naphthaleneacetic acid. and developed into viable plants. Key words: Pisum sativum L., regeneration, meristems, embryo axes, thidiazuron.

Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee
Keyword(s):  
Winter Wheat ◽  
Chinese Spring ◽  
Shoot Growth ◽  
Callus Formation ◽  
Triticum Aestivum L ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

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.

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