scholarly journals Organogenesis in eggplant (Solanum melongena L. cv. Embu) as affected by antibiotics and growth regulators

Author(s):  
E. A. de T. Picoli ◽  
P. R. Cecon ◽  
M. G. Fári ◽  
W. C. Otoni

The influence of antibiotics (cefotaxime, timentin, kanamycin and hygromycin) and growth regulators (indolacetic acid and 6-benzylaminopurine) was evaluated on eggplant organogenesis. Solanum melongena hypocotyl segments (6 to 10 mm length), taken from 16 to 20-days in vitro grown seedlings, were used as explants. The basic medium was composed by MS salts, Gamborg vitamins and 2% sucrose, solidified with agar 0.8% and pH adjusted to 5.7±0.2. Morphogenesis was impaired at 50 to 100 mg L-I kanamycin and 7.5 mg L-1 hygromycin. Both Timentin and cefotaxime reduced the frequency of regenerating explants meanwhile hyperhydricity was not affected. A decrease in root regeneration was observed with increasing cefotaxime concentrations, although, timentin had no effect on root regeneration, as compared to the control treatment. Interestingly, the number of adventitious roots was more noticeable at 0.25 mg L-I IAA plus 0.5 mg L-1 BAP. However, if just IAA was added led to higher number of regenerated roots compared to other treatments.

Growth regulators, phytohormones, both natural and artificial, are the main means to control plant ontogenesis. They are involved in regulating the processes of cell differentiation and cell divisions, the formation of tissues and organs, the changes in the rate of growth and development, the duration of the certain stages of ontogenesis. The main classes of phytohormones used in plant biotechnology, in particular, in the induction of haploid structures, are auxins and cytokinins. The mechanism of action of phytohormones on a cell is rather complicated and may have a different character. Understanding the characteristics of the action of phytohormones is complicated by the fact that the system of hormonal regulation of plant life is multicomponent. This is manifested in the fact that the same physiological process is most often influenced not by one, but by several phytohormones, covering a wide range of aspects of cell metabolism. In connection with the foregoing, the purpose of our work was to test a set of nutrient media with different basic composition and different proportions of phytohormones to determine the patterns of their influence on the processes of haploid structure induction in rape anther culture using accessions, developed at the Institute of Oilseed Crops NAAS. The material used was two accessions of winter rapeseed (No. 1 and No. 2) and one sample of spring rapeseed, provided by the Rapeseed Breeding laboratory of the Institute of Oilseed Crops. Incised inflorescences were kept against the background of low temperature of 6–8 ° C for several days, and then, under aseptic conditions, anthers with unripe pollen grains were isolated and planted on nutrient media differing in both basic mineral composition and content of phytohormones. MS (Murashige & Skoog 1962) and B5 (Gamborg et al 1968) media were used as basic media. Phytohormones were added to the basic media in various combinations – BA, 2,4-D, NAA at the concentrations of 0.1-0.6 mg/l. In each treatment up to 300 anthers were cultivated. Differences between treatments were evaluated using standard t-test. Studies have shown that in the anther culture of rapeseed on the tested nutrient media, morphogenic structures of different types (embryoids and callus) were originated. Synthetic auxin 2,4-D, regardless of the composition of the basic medium, caused the formation of structures of both types, though with a low frequency. Phytohormone BA of the cytokinin type had a similar effect. In this case, the frequency of structures was slightly higher, and the developed structures were represented mainly by embryoids. The joint action of cytokinin and auxin was the most favorable for the initiation of morphogenic structures. Such combination of phytohormones caused the formation of these structures with a frequency of 24.5-14.7% in the studied genotypes of winter rape. A similar effect of phytohormones on the induction and development of morphogenic structures was also observed in spring rape. In this case, a single basic MS medium was used. The experiment included treatments where phytohormones were absent (control), as well as various combinations of auxin and cytokinin. In the control treatment, the formation of new structures was not noted. In treatments with phytohormones, in addition to the medium with the combination of auxin and cytokinin, the medium in which only cytokinin was present was also rather effective. The treatment in which the action of auxin 2,4-D was combined with the action of another auxin, NAA, turned out to be practically ineffective. Thus, it was found that for the induction of morphogenic structures from microspores in rape anther culture of the tested genotypes, the combination of cytokinin with auxin, or the use of only single cytokinin BA without other phytohormones, had the most positive effect.


Author(s):  
A. Z. Revutskaya ◽  
A. V. Holubenko ◽  
N. V. Nuzhyna ◽  
H. O. Rudik ◽  
N. Yu. Taran

Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.


2015 ◽  
Vol 61 (3) ◽  
pp. 67-76
Author(s):  
Katarzyna Bączek ◽  
Izabela Szymborska-Sandhu ◽  
Anna Pawełczak ◽  
Zenon Węglarz

Summary An efficient method for in vitro propagation of bastard balm by enhanced axillary shoot branching has been developed. The material to establish in vitro culture was shoot tips collected from three-year-old plants in May. The shoots obtained from initial explants were placed on MS/B5 medium containing 0.1, 0.5 or 1.0 mg/l BA with 0.01 mg/l NAA or without the auxin. The highest number of shoots per explant was obtained on the medium with 1.0 mg/l BA (3.9 shoots per explant). For the rooting of shoots ½ MS/B5 with IBA (0.25, 0.50 and 1.0 mg/l) medium was used. The medium without plant growth regulators served as a control. The best root regeneration was observed on the medium without IBA (87.1% of cuttings rooted). IBA used in the medium for shoot rooting affected the morphological traits of obtained microcuttings but not affected their weight. Irrespective of auxin concentration in this medium, obtained microcuttings acclimated in ex vitro conditions very well.


2017 ◽  
Vol 48 (5) ◽  
Author(s):  
Khierallah & Al-Obaidy

This research was conducted in order to study the effect of explant type and some plant growth regulators on culture initiation of Stevia rebaudiana Bertoni in vitro. The experiments included surface sterilization and test two types of explants (shoot tips and stem nodes) and the impact of KIN and BA and IAA and IBA in the cultures initiation. Results revealed the efficiency of sodium hypochlorite (NaOCl) for disinfestation of explant at 0.050% concentration giving less contamination for shoot tips and stem nods (10% and 20% respectively). Results showed that shoot tips inoculated in MS medium plus KIN at 0.3 mg. L-1 was significantly increase the number of regenerated shoots as it produced 4.2 shoots per explant while medium without cytokinin (control) produced less number of shoots reached 1.4 shoots per explant. KIN treatment reduced shoots length as control treatment produced the highest length (6.74 cm).  The interaction between the explant type and BA concentration was significantly increase the number of regenerated shoots as shoot tips produced 3.6 shoots per explant in MS medium supplemented with 0.1 mg. L-1. BA treatment reduced shoots length as control treatment produced the highest length (6.74 cm). No positive effect was gain when auxins (IBA and IAA) were added in combination with cytokinin in culture medium. The above results can be adopted to established stevia in vitro culture successfully.


2015 ◽  
Vol 1 (2) ◽  
pp. 297-303 ◽  
Author(s):  
Sayeed Shahriyar ◽  
Soleh Akram ◽  
Koushik Khan ◽  
Md Faruk Miya ◽  
Md Abdur Rauf Sarkar

One of the goals of the experiment is to standardization of HgCl2 treatment for explants sterilization. The objectives also include developing a reproducible cost effective protocol for large scale production of Solanum tuberosum of Cardinal variety plantlets from selectively better clones through plant in vitro propagation methods. Selection of growth regulators for proper multiple shoots regeneration, elongation and root induction. To produce genetically uniform plantlets within a short time capable surviving in natural condition raised in in vitro environment. Shoot tip and nodal segment explants from field grown plants were used as experimental materials in this investigation. All explants were cultured on Murashige and Skoog medium supplemented with various plant growth regulators. For surface sterilization of explants, HgCl2 (0.1%) for 2 minutes was found to be most effective for complete destroying of surface pathogens and getting healthy tissues. Shoot regeneration was observed from both shoot tips and nodal explants for the studied plant. Maximum number of shoot per culture (17) was recorded and it also obtained the highest average length of the shoot (5cm) in Murashige and Skoog medium containing no hormone. On the other hand 6-benzyl amino purine (0.2mg/l) in 3 media showed the highest rate of shoot multiplication (73%) and the highest average length (4cm). In case of Gibberellic acid (0.1mg/l) in Murashige and Skoog media showed its highest rate of shoot regeneration (82%) and the highest average length (4.5cm). From the overall experiment it was observed that shoot tips are more responsive for micro propagation. In root induction Murashige and Skoog medium supplemented with different concentration (0.5, 1, 1.5 and 2mg/l) of indol-3-acetic acid and kinetin. Indol-3-acetic acid and kinetin (1.5+1.5 mg/l) showed its lowest rate of root regeneration (40%) and the average length of the root (1.5 cm). On the contrary Murashige and Skoog medium with no hormone showed the rate of root regeneration (96%) and the highest average length of the root (2.5 cm). The supplemented Murashige and Skoog media with no hormone showed the best performance for root regeneration.Asian J. Med. Biol. Res. June 2015, 1(2): 297-303


Author(s):  
Antonio Calabuig-Serna ◽  
Carolina Camacho-Fernández ◽  
Ricardo Mir ◽  
Rosa Porcel ◽  
Esther Carrera ◽  
...  

AbstractIn eggplant microspore embryogenesis, embryos are produced and then transformed into undifferentiated calli, instead of developing as true embryos. This is the main current bottleneck that precludes this process from being efficient. In this work we aimed to shed light on the factors involved in the successful in vitro development of eggplant haploid embryos by evaluating the role of growth regulators (GRs) in this process. We analyzed the endogenous levels of different GRs, including auxins, cytokinins and gibberelins, as well as salicylic, jasmonic and abscisic acid, in microspores and microspore-derived embryos at different culture stages. We also analyzed the same GR profiles in leaf and anther wall tissues of different eggplant backgrounds. Finally, we assessed the application of different GR combinations to the culture medium. Our results showed that in eggplant there are no genotype-specific endogenous GR profiles that can be associated to a high embryogenic response. Instead, the embryogenic response seems related to different GR accumulation patterns during in vitro culture. The changes observed in the endogenous levels of salicylic and abscisic acid were not related to the embryo transition. There were, however, changes in the levels of indole acetic acid and dihydrozeatin. The best GR combination to promote callus production was 0.5 mg/L 1-naphthaleneacetic acid (NAA) and 0.5 mg/L 6-benzylaminopurine (BAP). A 20% reduction of NAA and BAP reduced embryo production but produced structures more anatomically similar to embryos. These results shed light on the role of GRs during the development of microspore-derived embryos in eggplant microspore cultures.


2013 ◽  
Vol 7 (2) ◽  
Author(s):  
Ayad Assi Obaid ◽  
Wessam Malek Dawood ◽  
Nour Sabry Nasser

This study was conducted in plant tissue culture Lab. Horticulture Dept., College of Agriculture, Diyala University to evaluated the effect of plant growth regulators, North and South pole of magnetic field on common mandarin (Citrus reticulata L. Blanco) seedling growth and callus initiation. Seeds were cultured on solidified MS medium to study the impact of increasing magnetic field flood on the production of some plant growth regulators, alkaloids callus initiation. Results can be summarized as follows: Kinetin at concentrations (0.0, 1.0, 2.0, 3.0, 4.0) mg/L for 8 weeks of treatment showed that the control treatment gave taller roots (30) mm while the treatment 4.0 mg/L gave the highest number of shoots 1.7 shoot/ seedling. In the magnetic field experiment, the effect of north and south pole of magnetic field 200 mT for six periods (0 , 2 , 6, 11 , `14 , 18) days, showed that the treatment of 18 days gave the best shoot length 22mm. When the medium supplemented with 2mg/ l 2,4-D enhances callus initiation, while the combine of 2,4-D and TDZ reduced the rate of initiation and growth of callus. The production of hormones and alkaloids from mandarin callus exposed to1, 2 or3 magnets recorded the highest amount of Kin in the presence of 3 magnets giving 125.45 µg/ g while the highest amount of ABA, GA , IAA was found in control callus which gave (91.61, 76.25, 51.18)µg/ g respectively. Alkaloids yield showed the presence of Octapamine in the treatment of 3 magnets yielding 73.74 µg/g. The highest amount of Synephrine reached 366.99 µg/ g in the treatment of 3 magnets while the amount of Tyramine recorded 79.02 µg/ g in the treatment of two magnets. Ephedrine was higher in the callus that not being exposed reached 219.99 µg/ g.


Author(s):  
E. A. T. Picoli ◽  
W. C. Otoni ◽  
P. R. Cecon ◽  
M. Fári

The influence of increasing concentrations of naphthaleneacetic acid and the antibiotics cefotaxime, timentin, kanamycin, and hygromycin on eggplant (Solantun melongena L. cv. Embil) somatic embryogenesis was investigated. Cotyledon explants were excised from 16 to 20 days old in vitro grown seedlings. NAA promoted somatic embryogenesis, although its concentrations had no influence on the mean number of embryos. Callusing decreaSed significantly with increasing NAA concentrations. Morphogenesis was stopped with 50 to 100 mg L-1 kanamycin and 7.5 to 15 mg L-1 hygromycin. Although early globular embryos were observed up to 15 mg L-1, further embryo development was inhibited at 10 mg L-1. Interestingly, cefotaxime (250 and 500 mg L-1) promoted a marked effect on enhancing fresh weight of calli, accompanied by decrease in embryo regeneration, whereas timentin concentrations (150 and 300 mg L-1) did not affect embryo differentiation as compared to the control treatment.


2013 ◽  
Vol 38 (2) ◽  
pp. 65-87 ◽  
Author(s):  
Krystyna D. Kromer

The aim in view was investigation ofthe regenerative potential of rhizomes, bulbs and corms of ten monocotyledonous plant species from four families: <i>Amaryllidaceae</i> (<i>Haemanthus katharinae, Crinum abyssinicum, Leucojum vernum</i>), <i>Araceae</i> (<i>Spathiphyllum wallisii</i>), <i>Iridaceae</i> (<i>Crocus vernus, Iris germanica</i>), <i>Liliaceae</i> (<i>Hosta lancifolia, Muscari racemosum, Scilla laxiflora, Veltheimia viridifolia</i>) under conditions of in vitro culture. All the investigated species were capable of buld or, alternatively, bud and root regeneration. Different morphogenetic potential was noted between the particular families. A high regenerative potential under the conditions of culture applied was characteristic for plants of the <i>Liliaceae</i> and <i>Araceae</i> families, it was lower in plants belonging to <i>Amaryllidaceae</i> and lowest in those of the <i>Iridaceae</i> family. Plants from the <i>Liliaceae</i> family exhibited also the highest ability of callus formation, whereas <i>Amaryllidaceae</i> and <i>Iridaceae</i> plants possessed this ability in only a low degree. The influence of growth regulators of the auxin group (NAA, IAA, 2,4-D) and of cytokinins (K) on the initiation and course of organogenesis was tested. The results of the experiments indicate that auxins in interaction with kinetin gave the highest percentage of regenerating explants and also a large number of buds on the latter. Stimulation of callus tissue was highest under the influence of 2,4-D, and weaker when NAA IAA were used.


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