scholarly journals Micropropagation in vitro of essential oil rose hybrids obtained in embryoculture

2021 ◽  
Vol 38 ◽  
pp. 00139
Author(s):  
Natalia Yegorova ◽  
Irina Stavtzeva ◽  
Victor Zolotilov
Keyword(s):  
Essential Oil ◽  
Embryo Culture ◽  
Interspecific Hybrids ◽  
Culture Medium ◽  
High Variability ◽  
Ms Medium ◽  
Clonal Micropropagation ◽  
Hybrid Seedling ◽  
Stem Segments

The aim of the work was to study the features of clonal micropropagation of essential oil rose interspecific hybrids obtained in embryo culture in vitro. Analysis of 12 crossing combinations demonstrated that the frequency of hybrid seedling formation in the embryo culture varied from 0 to 71.4%. For clonal micropropagation obtained in vitro seedlings were divided into stem segments with a node and cultivated on MS culture medium supplemented with 0.5 mg/l BAP and 0.1 mg/l GA3. During the multiplication of 13 hybrids (R. alba × R. damascena cv. ‘Kazanlykskaya’) in 2-6 subcultures, high variability of the multiplication index (1.8-18.5 depending on the genotype and passage) was revealed. This parameter was maximum in the 3-4th subcultures. The best ability to micropropagation showed hybrid No. 37-14. Microshoots were rooted in vitro on ½ MS medium, containing for different hybrids 0.5 or 1.0 mg/l NAA; frequency – up to 80.5-100.0%. However, in No. 37-2, 37-19 and 37-31 on four tested media, the number of shoots with roots was only 0-35.4%.

scholarly journals Culture medium for mint micropropagation in vitro

2020 ◽  
Author(s):  
N.A. Yegorova ◽  
◽  
M.S. Zagorskaya ◽  
O.V. Yakimova ◽  
◽  
...  
Keyword(s):  
In Vitro Propagation ◽  
Culture Medium ◽  
Medium Composition ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Second Stage ◽  
Clonal Micropropagation ◽  
Propagation Technique

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.

scholarly journals Propagation of Sedum spectabile Boreau in Leaf Culture in Vitro

2012 ◽  
Vol 40 (1) ◽  
pp. 107 ◽  
Author(s):  
Cuiqin YANG ◽  
Yaoguo QIN ◽  
Xin SUN ◽  
Shu YUAN ◽  
Honghui LIN
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
Shoot Proliferation ◽  
Shoot Formation ◽  
Root Induction ◽  
Ms Medium ◽  
Shoot Induction ◽  
Leaf Base ◽  
Stem Segments

An efficient protocol was established for Sedum spectabile Boreau propagation. Various leaf parts were used as explants to regenerate plantlets, the stem segments of which were cultured for shoot proliferation and plantlet multiplication. The results showed that the leaf base was the optimal explant, as compared to both the middle and the top of leaves, for shoot formation. The highest shoot induction of 88.9% was observed on MS medium supplemented with 0.6 mg/l TDZ and 0.1 mg/l NAA. Hyperhydric leaves obtained in primary culture developed first into abnormal somatic embryos 10 days after subculture, and then into hyperhydric plantlets after an additional 10 days. The hyperhydric plantlets reversed to normal plantlets when plant growth regulators were removed from culture medium. Further, stem segments from reversed plantlets were used for shoot regeneration and root induction. Optimal shoot regeneration was obtained in MS medium containing 0.6 mg/l TDZ with 0.1 mg/l NAA. Root induction and root mean number were all higher on auxin-free medium than on medium containing auxins.

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

2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Survival Rate ◽  
Growth Regulators ◽  
In Vitro Propagation ◽  
Culture Medium ◽  
Culture Media ◽  
Ms Medium ◽  
Organic Additives ◽  
Clonal Micropropagation ◽  
Half Strength

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

scholarly journals Optimization of clonal micropropagation of Chrysanthemum

2019 ◽  
Vol 9 (4) ◽  
pp. 676-678
Author(s):  
I. D. Borodulina ◽  
A. Trufanova ◽  
G. Shevchenko ◽  
G. Sokolova ◽  
T. Plaksina
Keyword(s):  
Growth Regulators ◽  
Traditional Method ◽  
Proliferative Activity ◽  
Culture Medium ◽  
Ms Medium ◽  
Mineral Salts ◽  
Clonal Micropropagation ◽  
Murashige Skoog ◽  
High Proliferative Activity

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.

scholarly journals Clonal micropropagation of Thymus vulgaris L.

2020 ◽  
Vol 224 ◽  
pp. 04001
Author(s):  
A Sh Tevfik ◽  
N. A. Yegorova
Keyword(s):  
Plant Material ◽  
Culture Medium ◽  
Culture Media ◽  
Morphometric Parameters ◽  
Thymus Vulgaris ◽  
Ms Medium ◽  
Ex Vitro ◽  
Clonal Micropropagation ◽  
Propagation Stage

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).

scholarly journals GROWTH OF ‘PRATA-ANA’ BANANA’S MICROSHOOTS CLONE GORUTUBA FROM SYNTHETIC SEEDS:SUBSTRATES AND BAP CONCENTRATION

2017 ◽  
Vol 39 (5) ◽  
Author(s):  
JÚLIO CÉSAR GOMES PEREIRA ◽  
SELMA SILVA ROCHA ◽  
LUCIANA CARDOSO NOGUEIRA LONDE ◽  
MARCELA CAROLINE BATISTA DA MOTA ◽  
PABLO FERNANDO SANTOS ALVES ◽  
...  
Keyword(s):  
Culture Medium ◽  
Economic Importance ◽  
Leaf Length ◽  
Naphthalene Acetic Acid ◽  
Synthetic Seed ◽  
Ms Medium ◽  
World Production ◽  
Number Of Leaves ◽  
Alginate Matrix

ABSTRACT The banana crop stands out as an activity of great social and economic importance in Brazil, which occupies the fifth place in world production. Synthetic seed production is becoming promising for a micropropagation and in vitro conservation. The aim of the study was to analyze the conversion and growth of ‘Prata-anã’ banana’s microshoots clone Gorutuba from synthetic seed in MS medium and vermiculite, different substrates and concentrations of BAP (6-benzylaminopurine) associated with ANA (acetic naphthalene acid) in the constitution of its capsule were tested. The microshoots were immersed in the sodium alginate matrix (3%) and dripped in a solution of CaCl2.2H2O (100 mM) for complexation and then in KNO3 solution (100 mM) to decomplex. The experimental design was completely randomized in a 2 x 5 factorial design (substrate x BAP concentrations), containing different substrates (MS culture medium and vermiculite) and BAP concentrations (2.22, 4.44, 6.66, 8.88 and 13.32 µmol L-1) associated with NAA (naphthalene acetic acid) 0.54 µmol L-1, totaling 10 treatments, with 4 replicates, and that each replicate containing 5 seeds. The evaluations of conversion, number of leaves, leaf length, leaf height, number of roots, root length and oxidation were performed at 30 and 60 days.The use of the MS medium provided better growth results in relation to vermiculite as substrate, in which the different BAP concentrations did not differ from each other. It was found that, in MS culture medium, BAP concentrations above 8.88 µmol L-1 in the capsule composition are not indicated for microshoots growth.

scholarly journals In Vitro Seed Germination and Embryo Culture in Nothapodytes foetida (Wight) Sleumer

2015 ◽  
Vol 48 ◽  
pp. 23-31 ◽  
Author(s):  
S. Kaveri ◽  
Rao Srinath
Keyword(s):  
Seed Germination ◽  
Embryo Culture ◽  
Filter Paper ◽  
Agar Medium ◽  
Ms Medium ◽  
Mature Embryos ◽  
Nothapodytes Foetida ◽  
Half Strength ◽  
Paper Bridge

In vitro seed germination and embryo culture have been achieved in Nothapodytes foetida, this plant is known for its rich source of anticancer drug i. e., Camptothecin. In present study both normal and decoated seeds were subjected to different treatments viz., H2O, GA3, H2O2, H2SO4, chlorine water and mechanical scarification, further these were germinated on water agar medium (WA), filter paper bridge (FB), half strength MS (HMS) and full strength MS (FMS) medium. The highest percentage (69%) of germination was achieved from decoated seeds treated with 10mg/L GA3 and germinated on Filter Paper Bridge. And for embryo culture mature embryos were inoculated on MS medium containing various combination and concentrations of cytokinins (BAP, Kn and TDZ) and auxin (IAA and NAA) for rapid conversion into a plantlet. Among the different combinations of growth regulators; highest frequency (100%) of plantlet conversion was obtained on MS medium containing Kn (1.0mg/L) and NAA (0.2mg/L).

Effects of changing culture medium on preimplantation embryo development in rabbit

Zygote ◽  
2021 ◽  
pp. 1-6
Author(s):  
Haixia Wang ◽  
Wenbin Cao ◽  
Huizhong Hu ◽  
Chenglong Zhou ◽  
Ziyi Wang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Culture ◽  
Culture Medium ◽  
Cell Number ◽  
Apoptotic Index ◽  
Total Cell ◽  
Toxic Substances ◽  
Total Cell Number ◽  
Preimplantation Embryo Development

Summary Many studies have focused on the optimization of the composition of embryo culture medium; however, there are few studies involving the effect of a culture medium changing procedure on the preimplantation development of embryos. In this study, three groups were designed: a non-renewal group, a renewal group and a half-renewal group. The levels of reactive oxygen species (ROS), apoptotic index, blastocyst ratio and blastocyst total cell number were analyzed in each group. The results showed that the ROS level and the apoptotic index of blastocyst in the non-renewal group were significantly higher than in the renewal group and the half-renewal group (P < 0.05). The blastocyst ratio and blastocyst total cell number were significantly higher in the half-renewal group than that in non-renewal group and the renewal group (P < 0.05). These results demonstrated that the procedure of changing the culture medium influenced ROS level, apoptotic index, blastocyst ratio and total cell number of blastocysts. In addition, the result suggested that changing the culture medium may lead to a loss of important regulatory factors for embryos, while not changing the culture medium may lead to the accumulation of toxic substances. Half-renewal can alleviate the defects of both no renewal and renewal, and benefit embryo development. This study will be of high value as a reference for the optimization of embryo culture in vitro, and is very significant for assisted reproduction.

80 DEVELOPMENT OF A SYNTHETIC MEDIUM FOR THE IN VITRO CULTURE OF BOVINE EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 154 ◽  
Author(s):  
D. Moreno ◽  
A. Neira ◽  
L. Dubreil ◽  
L. Liegeois ◽  
S. Destrumelle ◽  
...  
Keyword(s):  
Growth Factors ◽  
Embryo Culture ◽  
Disease Transmission ◽  
Culture Medium ◽  
Culture Media ◽  
Synthetic Medium ◽  
Wilcoxon Test ◽  
Blastocyst Development ◽  
Negative Effect

In the majority of media for embryo culture, 2 of typical components used are FCS or BSA; however, the presence of FCS in the culture medium has been shown to have a negative effect on embryo quality and the use of animal-derived proteins in culture media increases the risks of disease transmission through in vitro embryo production. The aim of this study was to develop an in vitro embryo culture medium free from FCS and BSA, but with the addition of various growth factors and cytokines (GF-CYK: IGF-I, IGF-II, bFGF, LIF, GM-CSF) 50 ng mL–1 and (TGF-β1) 100 ng mL–1 supplemented with hyaluronan (HA) and recombinant albumin (RA). Bovine oocytes (n = 1043, 6 replicates) from abattoir ovaries were matured in TCM-199 medium with 60 μg mL–1 penicillin, 60 μg mL–1 streptomycin, and 10 ng mL–1 EGF for 24 h at 39°C and 5% CO2 in humidified air. Afterward, the oocytes were fertilized in IVF-TALP medium with 6 mg mL–1 fatty acid-free BSA and 1.7 IU mL–1 heparin for 18 h under the same conditions. After fertilization, presumptive zygotes were divided into two groups and cultured in 30 μL droplets of SOF supplemented with (1) 0.4% BSA + 5 μg mL–1 insulin, 5 μg mL–1 transferrin, and 5 ng mL–1 selenium (ITS) as a control; or (2) GF-CYK + 0.5 mg mL–1 HA + 0.15% RA (M1). Droplets were preserved under mineral oil in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 at 39°C. Blastocyst development and blastocyst diameter was observed at 7 and 8 days post-fertilization (dpf). Developmental and diameter data were analysed using the Wilcoxon test by using R software. The blastocyst rates were not significantly different between the control and M1 medium: at 7 dpf (22.9% ± 4.8 and 30.2% ± 3.0), and at 8 dpf (29.6% ± 5.1 and 37.4% ± 2.0 respectively; P > 0.05). The blastocyst diameter obtained with the M1 medium was significantly greater (P < 0.05) than that of the control at 7 dpf (173.3 μm ± 4.9 and 157.2 μm ± 4.1, respectively); however, no significant differences were observed at 8 dpf (190.3 μm ± 5.2 and 179.7 μm ± 5.3, respectively). In conclusion, the FCS- and BSA-free medium with GF-CYK, HA, and RA (M1) showed a comparable development rate to the control medium at 7 and 8 dpf. These growth factors and cytokines in association with hyaluronan and recombinant albumin have a synergistic action by promoting an increase in the blastocyst diameter at 7 dpf. This is fully synthetic method of embryo culture; it presents a valuable tool to reduce the risks of disease transmission via embryo transfer.

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