scholarly journals Clonal Micropropagation of representatives of the genus Dactylorhiza Neck. ex Nevski

2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES

2020 ◽  
Vol 23 (1) ◽  
pp. 178-190
Author(s):  
Jeillan Hussein ◽  
Diaa ibraheam

Marumi kumquat (Fortunella Japonica) is culture for its valuable nutritional value and medicinal importance in many regions of the world. The current study aimed to evaluate the effect of two types of media enriched with different concentrations of fructose and different plant growth regulators and different fructose concentration on in vitro propagation of Fortunella Japonica. The findings showed that the most effective treatment for explant surface sterilization was by using 0.1% HgCl2 for ten minutes which give best results for production contamination-free explants at the initiation cultures. At multiplication stage, WPM medium gave better results at all tested BA levels as compared with MS medium. No significant differences were showed by using BA alone or in combination with GA3 in the measured parameters. It has been observed that WPM medium supplemented with 0.5mgl-1 BA with the presence of 30mgl-1 fructose was able to give the highest shoot length (1.56cm) with maximum shoots number/explant 9.0 and highest leaves number/explant (21.0). The proliferated shoots were exposed to full strength MS medium salts supplemented with 2mgl-1 NAA which showed the highest ratio of rooting. In vitro rooted plantlets were gradually acclimatized and transferred to open air conditions, which recorded a high survive rate reached to 92%


Author(s):  
N.A. Yegorova ◽  
◽  
M.S. Zagorskaya ◽  
O.V. Yakimova ◽  
◽  
...  

The influence of the culture medium composition on the development of explants at the second stage of clonal micropropagation of mint (Mentha canadensis L. K59(4n)) was studied in order to improve the in vitro propagation technique. It was shown that the maximum multiplication rate (11.5) was provided by MS medium supplemented with BAP (1.0 mg/L), IAA (0.5 mg/L) and 2% sucrose.


2020 ◽  
Vol 30 (2) ◽  
pp. 243-252
Author(s):  
Ashok N Pyati

In vitro regenerative potential of protocorm like bodies (PLBs) of Dendrobium barbatulum were assessed. The formation of secondary PLBs from the primary PLBs that formed from the callus, it is derived from a few larger injured protocorm like bodies. The injured larger PLBs were cultured on half strength MS supplemented with different concentrations of BAP (0.32, 1.62, 3.23 and 16.15 μM), Zn (0.46, 2.28, 4.56 and 22.80 μM) and Kn (0.47, 2.33, 4.65 and 23.25 μM). The highest percentage of secondary PLBs (66.33%) were obtained on half strength MS medium supplemented with BAP 3.23 μM, after 5 weeks of culture. Then these clumps of PLBs were subcultured on half strength MS fortified with coconut water (CW), cane juice (CJ), peptone (P) and casein hydrolysate (CH) for regeneration. Among the complex organic additives 20% CW and 2.0 g/l P to half strength MS resulted in the development of plantlet 78.31 and 62.60%, respectively. Well developed plantlets were successfully acclimatized in the community pots having brick pieces, charcoal, decaying litter and coconut husk (1 : 1 : 1 : 1) gave maximum survival rate of 87.03%. Plant Tissue Cult. & Biotech. 30(2): 243-252, 2020 (December)


Author(s):  
Poornima Raj ◽  
J. Anbumalarmathi ◽  
S. Aruna Sharmili

An experiment was conducted for standardization of in vitro propagation technique of Bacopa monnieri (L.), a medicinal herb of India. Healthy leaf segments of the herb were used as explants with basic Murashige and Skoog (MS) medium containing various combinations of different growth regulators for callus, shoot and root initiation. The best callus induction percentage (95.47%) was observed on MS + 0.5 mg/L NAA and 2.0 mg/L BAP (T3). The maximum number of shoots (8), shoot length (9.30 cm) and shoot induction percentage (90.48%) was achieved on MS + 3.0 mg/L BAP and 1.0 mg/L Kn (ST4). The maximum number of roots (8) and root length (7) was observed on MS + 1.5 mg/L IAA (RT5). The rooted micro shoots were successfully hardened and acclimatized in green house and subsequently established in soil with survival rate of 90%.


2017 ◽  
Vol 66 (4) ◽  
pp. 544-548 ◽  
Author(s):  
Seir Antonio Salazar Mercado ◽  
Nelson Alfonso Vega Contreras

Cattleya trianae (Linden & Reichb.f., 1860), Colombian national flower, is in danger of extinction due to the destruction of its natural habitats and excessive collection for horticultural purposes. Therefore, in vitro culture is a tool for the conservation of threatened species. In this study we determined the most suitable culture medium for asimbytic seed germination and in vitro propagation of C. trianae. Initially, mature capsules were collected, the seeds were subsequently disinfected and seeded with the syringe method (Vendrame et al., 2007), to evaluate the effect of five media on the development of C. trianae after 20 weeks. The seedlings were transplanted and acclimated using different substrates. The best percentage (54.2%) of seedling formation after 20 weeks was found in MS + JP medium with significant differences (P <0.05: Tukey HSD). In this research, it is reported that the addition of organic additives to the MS medium improves the efficacy of this, and therefore, allows a greater growth and development of C. trianae under in vitro conditions.


2019 ◽  
Vol 9 (4) ◽  
pp. 676-678
Author(s):  
I. D. Borodulina ◽  
A. Trufanova ◽  
G. Shevchenko ◽  
G. Sokolova ◽  
T. Plaksina

Micropropagation of Chrysanthemum is an alternative to the traditional method of reproduction. Thanks to this method, the Chrysanthemum reproduction time is reduced to 3-4 months. For clonal micropropagation, sterile microshoots of Chrysanthemums of the varieties Snow White, Stranger, and Baltica White were used. At the stage of the micropropagation, the Murashige-Skoog (MS) medium with the full and half composition of mineral salts and growth regulators (kinetin, 6-benzylaminopurine, β-indolylacetic acid) were used. A universal culture medium for clonal micropropagation of all varieties of Chrysanthemum and optimal mediums, taking into account variety-specificity were established. It was noted that under in vitro conditions, high proliferative activity was observed in Neznakomka variety.


2020 ◽  
Vol 224 ◽  
pp. 04001
Author(s):  
A Sh Tevfik ◽  
N. A. Yegorova

Thymus vulgaris L. is one of the widely known spicy aromatic and medicinal plants. Thyme plant material is widely used in medicine, cooking and perfumery. To increase the efficiency of breeding and seed production, it is necessary to develop biotechnological techniques, in particular, clonal micropropagation. The aim of the research is to optimize the composition of culture media for the main stages of propagation in vitro and to select adaptation ex vitro conditions for the development of Thymus vulgaris. clonal micropropagation. The article presents the results of studies of explant morphometric parameters cultivated on 20 variants of culture media at firstsecond stages of micropropagation. It was found that the optimal culture medium at the introduction stage is MS medium with 1.0 mg/l Kin and 1.0 mg/l GA3, on which, on average, 2.2 microshoots per explant with a length of 1.9 cm were obtained. Both high vitrification rate of microshoots and formation of small shoots (0.6-0.9 cm) were observed on media supplemented with BAP or TDZ. The most effective culture medium at the proper propagation stage is MS with 1.0 mg/l Kin, on which 4.6 shoots per explant and the multiplication index 12.8 were obtained. It is advisable to root microshoots at the 3rd stage of micropropagation on MS culture medium supplemented with 1.0 mg/l IBA or 1.0 mg/l IAA. It has been shown that it is possible to obtain high plant survival rate (89.5%) during adaptation ex vitro, using a substrate consisting of peat and perlite (1:1).


2015 ◽  
Vol 22 (2) ◽  
pp. 69-73
Author(s):  
Kumud Saklani ◽  
Hem Pant ◽  
Vinod Bisht ◽  
Arun Singh ◽  
Vijay Rawat

The present study was conducted to improve upon the micropropagation protocol of Rose cultivars by modification of the MS medium through variations in sucrose and agar concentrations thereby enhancing the shoot multiplication and rooting efficiency of the nodal explants. High sucrose concentrations and low agar concentrations favoured shoot mutiplication during the in vitro stages. Enhanced multiplcation and growth was observed on sub culturing the mother explants with regenerated shoots on fresh MS medium containing sucrose (3.5%, w/v) and agar (0.6%, w/v), supplemented with lower concentrations cytokinin combination of BAP and KN (2.5 mgl-1+1.5 mgl-1) respectively. Half strength MS medium containing sucrose (2.0%, w/v) and agar (0.3%, w/v) with NAA and BAP (2.0 mgl-1+0.5 mgl-1) in combination was most effective for rooting.


Author(s):  
Rômulo Magno Oliveira Freitas ◽  
Narjara Walessa Nogueira ◽  
Sidney Carlos Praxedes

<p>O trabalho teve como objetivo desenvolver um protocolo de micropropagação de segmentos nodais de anador (<em>Justicia pectoralis</em>). Para isso foram realizados dois experimentos. O delineamento estatístico utilizado foi o inteiramente casualizado, com 15 repetições. Os segmentos de <em>J. pectoralis</em>, após desinfestados, foram cultivados em meio MS durante 30 dias. No primeiro experimento, esse material foi repicado em três meios de cultura (MS, WPM e B5) e após 77 dias foram avaliados comprimento de plântula, número de raízes, número de folhas e o número de segmentos nodais. Para o segundo experimento foram testadas duas citocininas (BAP e Cinetina) nas seguintes dosagens 0,0; 0,5; 5,0 e 20,0 mM. Aos 60 dias após a repicagem foram avaliadas as seguintes características: números de folhas, número de raízes e número de explantes por planta. O meio MS foi o que apresentou maior comprimento de plântula. As demais variáveis não diferiram entre os meios utilizados. Por isso o meio MS foi utilizado para o segundo experimento onde se verificou que a utilização de BAP proporcionou maior número de folhas e de explantes quando submetido à concentração de 20 mM. Dessa forma, para multiplicação de seg <em>Justicia pectoralis</em>, recomenda-se a utilização de meio MS com adição de 20mM de BAP.</p><p align="center"><strong><em>In vitro propagation of </em></strong><em>Justicia pectoralis<strong></strong></em></p><p><strong>Abstract</strong><strong>: </strong>The study aimed to establish a micropropagation protocol for <em>Justicia pectoralis</em> nodal segments. Two experiments were conducted. The statistical design was the completely randomized with 15 repetitions. After disinfestation, the segments of <em>J. pectoralis</em> were inoculated in the MS culture medium for 30 days. In the first experiment, the plant material was transferred to three culture media (MS, WPM and B5). The length of seedlings, number of roots, number of leaves, and number of nodal segments were evaluated at 77 days after transferring. In the second experiment two cytokinins (BAP and Kinetin) were tested in the following concentrations: 0.0; 0.5; 5.0 and 20.0 mM. At 60 days after transplanting the number of leaves, number of roots and number of explants per plant were evaluated. The MS medium induced the highest length of seedlings, but there was no effect for the other variables. Therefore, this medium was used for the second experiment, when it was found that BAP induced a larger number of leaves and explants when applied at 20 mM. Therefore, for multiplying <em>J. pectoralis</em> nodal segments we recommend the use of MS medium with 20 mM BAP.</p>


2013 ◽  
Vol 41 (2) ◽  
pp. 370 ◽  
Author(s):  
Marija MARKOVIĆ ◽  
Mihailo GRBIĆ ◽  
Matilda DJUKIĆ

During past decades, great attention has been paid to propagation of endangered plant taxa in order to preserve biodiversity. The aim of this study was to optimize a protocol for in vitro propagation of the critically endangered and decorative species Dianthus serotinus Waldst. et Kit. The effects of different concentration of MS salt (Murashige and Skoog) of the culture, medium pH and different carbohydrates (sucrose, glucose, and fructose) on shoot multiplication were examined. The best results were obtained on half-strength MS (Murashige and Skoog) medium, whose pH was 5.8, with sucrose supplied at a concentration of 3%, when shoots with 1-2 nodes or shoot tips (with terminal buds only) were used as explants. The shoots were rooted (76.7%) on half-strength MS medium containing 0.5 mg∙L-1 NAA (1-naphthaleneacetic acid). The obtained plantlets were successfully acclimatized (89%) in a 4:1 mixture of peat and sand and they flowered the following year. Presented protocol enables successful in vitro propagation of D. serotinus.


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