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

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.

Food Research ◽  
2020 ◽  
Vol 4 (S5) ◽  
pp. 115-123
Author(s):  
A.N. Salihan ◽  
N.A. Yusuf

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.


2021 ◽  
Vol 2 (5) ◽  
pp. 71-75
Author(s):  
Hasina Sultana ◽  
Lutfun Nahar ◽  
M. Mofazzal Hossain ◽  
Totan Kumar Ghosh ◽  
Md. Sanaullah Biswas

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.


2015 ◽  
Vol 21 (1) ◽  
Author(s):  
Esther Julia Naranjo ◽  
Osman Fernandez Betin ◽  
Aura Inés Urrea Trujillo ◽  
Ricardo Callejas Posada ◽  
Lucía Atehortúa Garcés

<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>


2021 ◽  
Vol 12 (4) ◽  
pp. 4731-4746

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.


2001 ◽  
Vol 7 (3-4) ◽  
Author(s):  
Zs. O. Kiss ◽  
A. Balogh ◽  
L. Fodorpataki

In vitro culture conditions were experimented for the relatively sensitive, but very esthaetic "Caribe" variety of carnation with uniformly dark violet flowers. Regeneration of new plants from shoot apex meristems can be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. Callus formation as a prerequisite for the induction of somaclonal variability can be achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Acclimation of the obtained mericlones to the ex vitro conditions was also evaluated.  


2003 ◽  
Vol 9 (3-4) ◽  
Author(s):  
Zs. O. Kiss ◽  
A. Mándy

Callus formation, as a prerequisite for the induction of somaclonal variability, was achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Regeneration of new plants from shoot apex meristems could be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. These in vitro treatments may contribute to a more efficient micropropagation of the Rimini variety of carnation.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva

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.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig

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.


Plants ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1261
Author(s):  
Robin Lardon ◽  
Danny Geelen

Plant regeneration is essential for survival upon wounding and is, hence, considered to be a strong natural selective trait. The capacity of plant tissues to regenerate in vitro, however, varies substantially between and within species and depends on the applied incubation conditions. Insight into the genetic factors underlying this variation may help to improve numerous biotechnological applications that exploit in vitro regeneration. Here, we review the state of the art on the molecular framework of de novo shoot organogenesis from root explants in Arabidopsis, which is a complex process controlled by multiple quantitative trait loci of various effect sizes. Two types of factors are distinguished that contribute to natural regenerative variation: master regulators that are conserved in all experimental systems (e.g., WUSCHEL and related homeobox genes) and conditional regulators whose relative role depends on the explant and the incubation settings. We further elaborate on epigenetic variation and protocol variables that likely contribute to differential explant responsivity within species and conclude that in vitro shoot organogenesis occurs at the intersection between (epi) genetics, endogenous hormone levels, and environmental influences.


Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Sara Rostampour ◽  
Haleh Sohi ◽  
Ali Dehestani

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.


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