scholarly journals EFFECT OF GENOTYPE ON THE in vitro REGENERATION OF Stevia rebaudiana VIA SOMATIC EMBRYOGENESIS

2015 ◽  
Vol 21 (1) ◽  
Author(s):  
Esther Julia Naranjo ◽  
Osman Fernandez Betin ◽  
Aura Inés Urrea Trujillo ◽  
Ricardo Callejas Posada ◽  
Lucía Atehortúa Garcés
Keyword(s):  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Stevia Rebaudiana ◽  
Dichlorophenoxyacetic Acid ◽  
Embryogenic Calli ◽  
Medicinal Properties ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Diabetes And Obesity ◽  
Embryogenic Response

<p class="p1"><strong>ABSTRACT</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae) is a plant of economic importance because of its medicinal properties and the presence of sweetener compounds on its leaves. These compounds can be a substitute for sucrose in a wide variety of products used by persons with diabetes and obesity problems. To standardize an efficient and effective propagation method for the different Stevia genotypes grown in Colombia, this study evaluated the effect of different combinations of the plant growth regulators 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 6-(gamma, gamma-dimethylallylamino) purine (2iP) and Zeatin on the induction and development of somatic embryos. Adenine and coconut water were also evaluated as supplements in the basal culture medium Murashige and Skoog Basal Salt Mixture (MS) with glutamine. The combination of 2,4-D (18.09 μM) and 2iP (7.38 μM) produced the highest number of somatic embryos per explant, which had well-defined characteristics. The genotype showed a significant effect on the embryogenic response. In the “SRQ-93” genotype, the formation and development of somatic embryos was achieved, whereas the genotypes “Bertoni” and “Morita II” only yielded embryogenic and non-embryogenic calli, respectively. The conversion to seedlings was achieved on the regeneration medium containing gibberellic acid (GA<span class="s1">3</span>) (0.29 μM) and activated charcoal. </p><p class="p1"><strong>RESUMEN</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae), es una planta de gran importancia económica debido a sus propiedades medicinales y a la presencia de compuestos endulzantes en sus hojas, los cuales pueden sustituir la sacarosa en gran variedad de productos utilizados por personas con problemas de diabetes y obesidad. Con el propósito de estandarizar un método de propagación eficiente y efectivo para diferentes genotipos de Stevia cultivados en Colombia, en la presente investigación se evaluó el efecto sobre la inducción y desarrollo de embriones somáticos de diferentes combinaciones de los reguladores de crecimiento vegetal 2,4-D, IAA, IBA, 2iP y Zeatina, además de los suplementos adenina y agua de coco en el medio de cultivo basal Murashige y Skoog (1962), adicionado con glutamina. Con la combinación 2,4-D (18.09 μM) y 2iP (7.38 μM) se obtuvo el mayor número embriones somáticos por explante con características bien definidas. El genotipo tuvo un efecto significativo en la repuesta embriogénica, en el genotipo “SRQ-93” se logró la formación y el desarrollo de embriones somáticos, mientras que en los genotipos “Bertoni” y “Morita II”, solo se obtuvo callo embriogénico y no embriogénico respectivamente. La conversión a plántulas se alcanzó en el medio de regeneración conteniendo GA3 (0.29 μM) y carbón activado.</p><p class="p2"> </p>

scholarly journals High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

2014 ◽  
Vol 6 (1) ◽  
pp. 85-91
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Devi Sunitibala HUIDROM
Keyword(s):  
High Frequency ◽  
Somatic Embryos ◽  
Cell Layer ◽  
Sucrose Concentration ◽  
Dichlorophenoxyacetic Acid ◽  
Thin Cell Layer ◽  
Regeneration System ◽  
Transverse Thin Cell Layer ◽  
2,4 Dichlorophenoxyacetic Acid

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

scholarly journals In vitro regeneration of groundnut: Changes in antioxidative enzymes and histological studies

2014 ◽  
Vol 59 (1) ◽  
pp. 63-73 ◽  
Author(s):  
Rohini Trivedi
Keyword(s):  
Antioxidative Enzymes ◽  
Stress Responses ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Stage Iv ◽  
Dichlorophenoxyacetic Acid ◽  
In Vitro Morphogenesis ◽  
Histological Studies ◽  
2,4 Dichlorophenoxyacetic Acid

Reactive oxygen species (ROS) produced during stress responses are implicated in a number of cellular responses including morphogenesis. The present study was undertaken to study the changes in antioxidative enzymes during in vitro morphogenesis of groundnut from de-embryonated cotyledon explants cultured on Murashige and Skoog?s medium supplemented with 5.0 mg l-1 benzyl-adenine and 2.0 mg l-1 2,4-dichlorophenoxyacetic acid. During the early in vitro ontogenic stages of groundnut, the activity of peroxidase (POD) and polyphenol oxidase (PPO) increased from stage 0 (0 day) to stage II (14 days) and decreased during stage III (25 days) and stage IV (45 days). The activity of superoxide dismutase (SOD) showed an inverse trend. The results could be correlated with the acquisition of competence, de-differentiation, division and induction which occurred during shoot organogenesis. Histological studies also showed that the mode of in vitro morphogenesis from the groundnut explants was via shoot organogenesis. In light of the above study, it could be concluded that the change in activity of the antioxidative enzymes studied could be used as a marker to characterize the mode of plant regeneration.

Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus

2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Symptomatic Treatment ◽  
Dichlorophenoxyacetic Acid ◽  
Leucojum Aestivum ◽  
Anisic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

scholarly journals Somatic Embryogenesis In Rosa Chinensis CV. ‘Parson’s Pink China’

2022 ◽  
Author(s):  
Yanfang Cai ◽  
Lintao Tang ◽  
Haixia Chen ◽  
Yufan Li ◽  
Rong Liu ◽  
...  
Keyword(s):  
Somatic Embryo ◽  
Somatic Embryos ◽  
Germination Rate ◽  
Survival Rates ◽  
Dichlorophenoxyacetic Acid ◽  
Embryogenic Calli ◽  
Rosa Chinensis ◽  
Induction Rate ◽  
Secondary Embryos ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract This study investigated the effects of explant type, plant growth regulator concentration, callis status, medium conversion time, and medium tilt on the growth of rose somatic embryos. The results showed that Rosa chinensis cv. ‘Parson’s Pink China’ leaves could induce normal embryogenic calli but petioles could not. When the 2,4-dichlorophenoxyacetic acid concentration was 3.0 mg/L, the callis induction rate was the highest in the embryo proliferation medium (EP medium) supplemented with 0.5mg/L kinetin, and white and reddish-brown translucent calli were the main type of embryogenic calli induced. As the culture time in EP medium was extended, the relative induction rate for secondary embryos and multicotyledon secondary embryos gradually increased when transferred to embryo maturation medium (EM medium), but the induction rate for somatic embryos decreased. Placing the EM medium at an angle of 45° made the somatic embryos germinate faster and the germination rate was also higher. The germination buds produced by the somatic embryos with two cotyledons showed the fastest germination and greatest survival rates. The results of this experiment will help improve somatic embryo regeneration rates and explore new ways of regeneration, and lays the foundation for further optimization of the somatic embryo genetic transformation system.

scholarly journals Plant regeneration from protoplasts of Gentiana straminea Maxim

2016 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Guomin Shi ◽  
Lina Yang ◽  
Tao He
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Protoplast Culture ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Embryogenic Calli ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Solid Liquid

AbstractA protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

scholarly journals In vitro regeneration of Didymopanax morototoni

2006 ◽  
Vol 66 (2a) ◽  
pp. 455-462 ◽  
Author(s):  
E. T. H. Franco ◽  
L. B. Gavioli ◽  
A. G. Ferreira
Keyword(s):  
Plant Regeneration ◽  
Somatic Embryos ◽  
Woody Plant ◽  
In Vitro Regeneration ◽  
Casein Hydrolysate ◽  
Forest Species ◽  
Embryogenic Calli ◽  
Woody Plant Medium ◽  
Germinated Seeds

The present study aimed at establishing a complete plant regeneration protocol for Didymopanax morototoni (matchwood), a native Brazilian forest species. Four types of explants (root, shoot, node, and cotyledonary leaves) were obtained from in vitro germinated seeds. In the first step, woody plant medium (WPM) with casein hydrolysate (250 mgL-1 ) and 2,4-D (1.0 and 5.0 mgL-1) were used combined with kinetin (0.1 and 1.0 mgL-1). Twenty days after inoculation, the material was evaluated. Embryogenic calli were split, transferred to expression medium with several combinations of NAA and KIN, and moved to fresh medium after 60 days. Light did not interfere in embryo expression. Somatic embryos were formed either from individual cells or cell clusters. Plantlets were obtained in WPM medium and 10 gL-1 of sucrose with no plant regulator, or using 0.1 mgL-1 BAP and 0.5 mgL-1GA. Plantlets from somatic embryos of D. morototoni developed in 33% of the cases.

scholarly journals Somatic Embryogenesis in Four Tree Legumes

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Premananda Das
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Friable Embryogenic Callus ◽  
Napthaleneacetic Acid ◽  
Acacia Catechu ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis was achieved in four leguminous tree species, that is, Acacia catechu, Acacia arabica, Hardwickia binata, and Dalbergia sissoo using immature zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 0.25–1.0 mg/l Kn (kinetin) and 2.0–3.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) or NAA (1-napthaleneacetic acid) and 3% sucrose. MS medium containing 2.0 mg/l 2,4-D and 1.0–1.5 mg/l Kn was noted to be most effective in inducing friable embryogenic callus (FEC); the number of somatic embryos per culture varied in MS medium supplemented with 1.0–2.0 mg/l 2,4-D or NAA and 0.25–1.5 mg/l kinetin. The maximum number of somatic embryos was obtained in MS medium containing 1.5–2.0 mg/l 2,4-D or NAA and 1.0–1.5 mg/l kinetin; proliferation of embryogenic calli was enhanced in cultures having 1.0–2.0 mg/l 2,4-D, 1.0–1.5 mg/l kinetin, and 400–600 mg/l L-Proline. The somatic embryos in various shapes and sizes after the first subculture on MS medium supplemented with 0.1 mg/l IAA and 0.25 mg/l BA; developed shoots and rooted in strength MS medium supplemented with 0.1 mg/l IBA or IAA. The somatic embryo-derived plantlets were transferred to the field after being hardened in the climate-controlled hardening chamber.

scholarly journals High frequency callus induction and proliferation of MD2 pineapple (Ananas comosus)

Food Research ◽  
2020 ◽  
Vol 4 (S5) ◽  
pp. 115-123
Author(s):  
A.N. Salihan ◽  
N.A. Yusuf
Keyword(s):  
Callus Culture ◽  
Callus Induction ◽  
Callus Formation ◽  
In Vitro Regeneration ◽  
Ananas Comosus ◽  
Dichlorophenoxyacetic Acid ◽  
Fresh Weight ◽  
Metabolites Production ◽  
2,4 Dichlorophenoxyacetic Acid

Ananas comosus var. MD2 is currently the most preferred pineapple variety in the international market due to its pleasant aroma and high Brix acidity ratio. In vitro approaches such as callus culture is promising in producing disease-free plantlet. However, there are limited studies reported on callus culture of MD2 variety despite the potential of in vitro regeneration through biotechnological advances. The purpose of the study was to determine the effect of plant growth regulators (PGRs) i.e., 2,4- dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP) and Thidiazuron (TDZ) on callus induction from leaf explant of MD2 pineapple. Leaf base explants were cultured on Murashige and Skoog (MS) media supplemented with varying concentration of 2,4-D (0.5 to 6.0 mg/L) alone and in combination with BAP (1.0 to 3.0 mg/L). The frequency of callus induction was seen significantly highest (91.67±8.33%) with maximum callus fresh weight (0.25±0.07 g) at a combination of 2.0 mg/L 2,4-D and 2.0 mg/L BAP. The shortest duration of callus formation was seen on day 12 with the lowest concentration of 2,4-D, 0.5 mg/L. There is a moderate correlation between the earliness of callus formation and the frequency of callus induction (P<0.01). The most favourable media for callus proliferation was 6.0 mg/L 2,4-D and 2.0 mg/L TDZ as the highest fresh weight of 1.52±0.03 g was recorded. Callus culture has the potential to be a source of plant material and secondary metabolites production. In this study, 2,4-D and BAP have successfully induced callus in MD2 pineapple.

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 The miR396–GRF Regulatory Module Controls the Embryogenic Response in Arabidopsis via an Auxin-Related Pathway

2019 ◽  
Vol 20 (20) ◽  
pp. 5221 ◽  
Author(s):  
Szczygieł-Sommer ◽  
Gaj
Keyword(s):  
Positive Control ◽  
Dichlorophenoxyacetic Acid ◽  
Regulatory Relationship ◽  
Gus Reporter ◽  
Regulatory Module ◽  
Genes Encoding ◽  
Cell Transcriptome ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

In plants, microRNAs have been indicated to control various developmental processes, including somatic embryogenesis (SE), which is triggered in the in vitro cultured somatic cells of plants. Although a transcriptomic analysis has indicated that numerous MIRNAs are differentially expressed in the SE of different plants, the role of specific miRNAs in the embryogenic reprogramming of the somatic cell transcriptome is still poorly understood. In this study, we focused on performing a functional analysis of miR396 in SE given that the transcripts of MIR396 genes and the mature molecules of miR396 were found to be increased during an SE culture of Arabidopsis [1]. In terms of miR396 in embryogenic induction, we observed the SE-associated expression pattern of MIR396b in explants of the β-glucuronidase (GUS) reporter line. In order to gain insight into the miR396-controlled mechanism that is involved in SE induction, the embryogenic response of mir396 mutants and the 35S:MIR396b overexpressor line to media with different 2,4-Dichlorophenoxyacetic acid (2,4-D) concentrations was evaluated. The results suggested that miR396 might contribute to SE induction by controlling the sensitivity of tissues to auxin treatment. Within the targets of miR396 that are associated with SE induction, we identified genes encoding the GROWTH-REGULATING FACTOR (GRF) transcription factors, including GRF1, GRF4, GRF7, GRF8, and GRF9. Moreover, the study suggested a regulatory relationship between miR396, GRF, and the PLETHORA (PLT1 and PLT2) genes during SE induction. A complex regulatory relationship within the miR396–GRF1/4/8/9–PLT1/2 module that involves the negative and positive control of GRFs and PLT (respectively) by miR396 might be assumed.

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