scholarly journals Efficiency of in vitro culture techniques applied to soybean (Glycine max (L.) Merr.) accessions from the VIR collection

2021 ◽  
Vol 182 (4) ◽  
pp. 137-142
Author(s):  
E. S. Korshikova ◽  
K. M. Ershova ◽  
Yu. A. Moksheninova ◽  
Yu. V. Ukhatova
Keyword(s):  
Callus Formation ◽  
Research Trend ◽  
Culture Techniques ◽  
New Genes ◽  
Wide Range ◽  
Ability To Regenerate ◽  
Presidential Decree ◽  
Genetic Technologies ◽  
Adverse Environmental Conditions

Using a wide range of modern biotechnologies and genetic techniques to study plant germplasm accessions held by VIR makes it possible to procure valuable results, required for the development of new high-yielding cultivars adapted to adverse environmental conditions and possessing specified technological properties, particularly to identify and mark new genes and alleles useful for plant breeding. This research trend is in line with Presidential Decree No. 680 “Concerning the development of genetic technologies in the Russian Federation”. Soybean is among the key crops in agricultural production, but the use of next-generation breeding tools to obtain new soybean cultivars with desired properties is still limited. Successful application of novel methods also requires new approaches to studying soybean accessions, specifically their ability to regenerate and produce calluses for subsequent inclusion in biotechnological programs.Ten soybean accessions of various origin, contrasting in ripening schedules, were selected to study the possibility of effective introduction into in vitro culture and further assessment of their ability to produce calluses and regenerate in in vitro culture. The work included evaluating the effects of different seed sterilization techniques (one-step sterilization, using a commercial bleach, and two-step one, combining the impacts of a chlorine-containing preparation and hydrogen peroxide), types of explants (epicotyls, hypocotyls, cotyledon nodes, and cotyledon leaf segments), and phytohormone composition of nutrient medium: (1) MS + 1.13 mg/L BAP + 0.5 mg/L HA, and (2) MS +1 mg/L BAP + 0.1 mg/L IAA).The assessment results showed that the option of two-step seed sterilization was the most effective for soybean at the stage of in vitro culture initiation, while hypocotyls, epicotyls, and cotyledon nodes had the highest callus formation ability in both types of nutrient media.

scholarly journals Immature embryo is the potential source for in vitro plant regeneration in Jatropha curcas

2014 ◽  
Vol 20 ◽  
pp. 125-134
Author(s):  
MR Islam ◽  
MA Bari
Keyword(s):  
Developmental Stages ◽  
Callus Formation ◽  
Immature Embryo ◽  
Basal Medium ◽  
Coconut Water ◽  
Immature Embryos ◽  
Plantlet Formation ◽  
Potential Source ◽  
Wide Range

Context: Jatropha belongs the spurge family Euphorbiaceae. Special interest mounting for its biodiesel which has created enthusiasm in cultivation of the species for oil extraction. Objectives: The study was conducted to develop the protocol for tissue and callus culture in Bangladeshi Jatropha curcus plant particularly to identify the most suitable explants for its wide scale micropropagation. Materials and Methods: Immature embryos taken from four developmental stages of fruits were cultured on growth regulator free MS liquid medium. After fifteen days of germination, elongated hypocotyls and two cotyledonary leaves were used as explants. Results: Embryo derived seedlings acted as the potential source of explants both for callus and plantlets. The immature embryo of size 0.87cm produced highest callus formation (83.33%) on MS medium supplemented with lower concentration of 2, 4-D (0.5 mg/l) and coconut water 2% (v/v). Immature embryos grown on MS basal medium supplemented with 2,4-D (0.2 mg/l, 0.5 mg/l and 1.0 mg/l) alone or in combination with coconut water 2% (v/v) exhibited a wide range of callus induction percentage (26-100%) for hypocotyls and (20 - 40%) for cotyledonary leaves. Conclusion: The age of immature embryo and addition of growth adjuvants and growth additive to the culture medium played the role in promoting better callus and plantlet formation. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17726 J. bio-sci.  20:  125-134, 2012

scholarly journals New human chromosomal safe harbor sites for genome engineering with CRISPR/Cas9, TAL effector and homing endonucleases

2018 ◽  
Author(s):  
Stefan Pellenz ◽  
Michael Phelps ◽  
Weiliang Tang ◽  
Blake T. Hovde ◽  
Ryan B. Sinit ◽  
...  
Keyword(s):  
Genome Engineering ◽  
Fluorescent Proteins ◽  
Population Level ◽  
Safe Harbor ◽  
Homing Endonucleases ◽  
Transgene Integration ◽  
New Genes ◽  
Wide Range

AbstractSafe Harbor Sites (SHS) are genomic locations where new genes or genetic elements can be introduced without disrupting the expression or regulation of adjacent genes. We have identified 35 potential new human SHS in order to substantially expand SHS options beyond the three widely used canonical human SHS,AAVS1, CCR5andhROSA26. All 35 potential new human SHS and the three canonical sites were assessed for SHS potential using 9 different criteria weighted to emphasize safety that were broader and more genomics-based than previous efforts to assess SHS potential. We then systematically compared and rank-ordered our 35 new sites and the widely used humanAAVS1, hROSA26andCCR5sites, then experimentally validated a subset of the highly ranked new SHS together versus the canonicalAAVS1site. These characterizations includedin vitroandin vivocleavage-sensitivity tests; the assessment of population-level sequence variants that might confound SHS targeting or use for genome engineering; homology–dependent and –independent, SHS-targeted transgene integration in different human cell lines; and comparative transgene integration efficiencies at two new SHS versus the canonicalAAVS1site. Stable expression and function of new SHS-integrated transgenes were demonstrated for transgene-encoded fluorescent proteins, selection cassettes and Cas9 variants including a transcription transactivator protein that were shown to drive large deletions in aPAX3/FOXO1fusion oncogene and induce expression of theMYF5gene that is normally silent in human rhabdomyosarcoma cells. We also developed a SHS genome engineering ‘toolkit’ to enable facile use of the most extensively characterized of our new human SHS located on chromosome 4p. We anticipate our newly identified human SHS, located on 16 chromosomes including both arms of the human X chromosome, will be useful in enabling a wide range of basic and more clinically-oriented human gene editing and engineering.

scholarly journals Stimulation of Stem Callus and Regeneration of Ficus elastica "Decora" and from Apical Tips in vitro

2013 ◽  
Vol 7 (2) ◽  
pp. 5-12
Author(s):  
Ragad M. Abdullah ◽  
Mazahim K. AL-Mallah
Keyword(s):  
Callus Formation ◽  
Potassium Nitrate ◽  
Shoot Tips ◽  
Peat Moss ◽  
Ms Medium ◽  
Ability To Regenerate ◽  
Calli Cultures ◽  
Clear Increase ◽  
Stimulation Of

Calli cultures of stems and leaves explants excised from field -grown rubber, Ficus elastica Decora, plants were formed on agar-solidified Murashige and Skoog (MS) medium.The results proved that MS medium supplemented with 1.0 mg L-1 benzyl adenin (BA) and 0.8mg L-1 2,4-dichloro-phenoxy acetic acid (2,4-D) was suitable to stimulate stem’s callus at ratio 87.5% . Whereas supplementation of MS medium with 0.5 mg L-1 of both BA and Indole -3-butyric acid (IBA)encouraged callus formation to reach76.6%. Leaves showed responses for callus initiation up to 75% at the same media. Stem calli showed limited ability to regenerate shoots on both agar solidified MS medium containing 1.0 mg L-1 2,4-D and kinetin (kin) 0.5 mg L-1 and MS medium supplemented with 0.5 mg L-1 of both BA and IBA. Transferring of shoots to differentiation medium (MS+0.5 mg L-1 BA+ 0.1 mg L-1 IBA) stimulated the growth and elongation of these shoots. Shoots were transferred to agar-solidified MS medium free from growth regulators failed to form roots. Because of the less number of shoots, other rooting media were not tested. The data showed, that shoot tips succeeded to regenerate shoots when they were cultured in different MS. The results proved clear increase in chlorophyll and protein content of the rubber shoots as compared with content of field grown rubber plants. It was noticed that agar-solidified MS medium supplemented with 4.0 mg L-1 BA was considered the optimum medium for shoots regeneration. All plants regenerated from shoot tips were readily rooted in agar-solidified MS medium with increasing Potassium Nitrate KNO3 from 1900 mg L-1 to 2000 mg L-1, and at the same medium supplemented with 3.0 mg L-1 IBA and 1.0 mg L-1 BA. All these plants were successfully acclimated and transferred to peat moss.

scholarly journals PENGGUNAAN 2,4-D UNTUK INDUKSI KALUS KLON KAKAO UNGGUL SULAWESI 1

2020 ◽  
Vol 2 (2) ◽  
Author(s):  
Yulianti Rasud ◽  
Moh. Habil ◽  
Tony Tony
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
Culture Techniques ◽  
Randomized Design ◽  
Honestly Significant Difference ◽  
Significant Difference ◽  
Completely Randomized Design ◽  
Difference Test ◽  
Honestly Significant Difference Test

ABSTRACT The multiplication of cocoa clones in conventional Sulawesi has not yet been able to fulfill the demand for large quantities of seeds because it is limited by the number of shoots and branches ready to be tapped, connected and oculated and takes longer to produce large quantities of seeds. One alternative in overcoming this problem is plant proragation using tussue culture techniques.  The aim of this experiment was to determine the appropriate of 2,4-D for callus induction of superior cocoa clones Sulawesi via in vitro culture.  This experiment used Completely Randomized Design with five treatments, namely 0.50 ppm 2,4-D, 0.75 ppm 2,4-D, 1.00 ppm 2,4-D, 1.25 ppm 2,4-D and 1.50 ppm 2,4-D.  Parameters observed consisted of the time, percentage, color and texture of calli.  Data was analized by using analysis of variance and differences between mean treatments were determined by Honestly Significant Difference Test at 5% level.  Results of this experiment indicated that the ability of different callus induction at various concentrations of 2,4-D for superior cocoa clones in Sulawesi 1 was tried.  it was obtained the quickest callus formation at concentration 0.50ppm 2,4-D namely average 4.22 WAC with the percentage of callus formation was up to 99,33%. Keywords: Callus Induction, Clones Sulawesi 1, 2,4-D ABSTRAK Perbanyakan klon kakao Sulawesi secara konvensional saat ini belum dapat memenuhi permintaan bibit dalam jumlah besar karena sangat dibatasi oleh jumlah tunas dan cabang yang siap disetek, disambung, dan diokulasi serta dibutuhkan waktu yang lebih lama untuk menghasilkan bibit dalam jumlah besar. Salah satu alternatif dalam mengatasi masalah tersebut adalah perbanyakan tanaman dengan menggunakan teknik kultur jaringan.  Penelitian ini bertujuan untuk memperoleh protokol yang tepat dalam menginduksi kalus sebagai upaya awal dalam perbanyakan tanaman kakao melalui embryogenesis. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan 5 level perlakuan yaitu 0,50 ppm 2,4-D, 0,75 ppm 2,4-D, 1,00 ppm 2,4-D, 1,25 ppm 2,4-D dan 1,50 ppm 2,4-D. Pengamatan dilakukan terhadap saat muncul kalus, persentase eksplan berkalus, warna kalus dan tekstur kalus.  Data diolah dengan analisis ragam dan perbedaan antar perlakuan ditentukan dengan Uji Beda Nyata Jujur pada taraf 5%. Hasil penelitian menunjukkan kemampuan induksi kalus berbeda pada berbagai konsentrasi 2,4-D untuk klon kakao unggul Sulawesi 1 yang dicobakan. Saat muncul kalus paling cepat diperoleh pada konsentrasi 0,5 ppm 2,4-D yaitu rata-rata 16,67 HST dengan persentase pembentukan kalus tertinggi mencapai 99,33%.  Selanjutnya, warna dan tekstur kalus yang dihasilkan yaitu remah putih dan remah kecoklatan. Kata Kunci: Induksi Kalus, Klon Sulawesi 1, 2,4-D.

scholarly journals Stabilization of soft wheat selection material which is the result of crossing with wild relatives

2021 ◽  
Vol 28 ◽  
pp. 83-87
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
T. P. Nargan ◽  
M. S. Chekalova
Keyword(s):  
Winter Wheat ◽  
Anther Culture ◽  
Callus Formation ◽  
Wild Relatives ◽  
Soft Wheat ◽  
Ability To Regenerate ◽  
Androgenesis In Vitro ◽  
Callus Regeneration ◽  
Soft Winter Wheat

Aim. Testing the haploproduction ability of 30 hybrids of winter soft wheat. Methods. In vitro culture of isolated anthers of wheat. The percentage of callus and regeneration of green plants for each genotype calculated as a percentage of the planted anthers. Results. The differences in the frequency of callus induction and the ability to regenerate plants in the process of androgenesiss in vitro of winter soft wheat were detected. The microspores of 17 of 30 hybrids formed callus by in vitro anther culture were shown. The intensity of one process was different: more than half of the genotypes (18 pcs.) were characterized by a low percentage of callus (from 0.10 to 1.0%), 6 genotypes - medium (from 1.0 to 3.0%), and three - high (4.36%; 15.11% and 15.81%, respectively). Conclusions. Genotype-specific of microspores morphogenetic reactions of soft winter wheat in the process of androgenesis in vitro were revealed Samples P26 and P27 showed the highest level of callus formation. The 10 green regenerating plants were obtained. Keywords: hybrids, soft winter wheat, anther culture in vitro, callus, regeneration.

scholarly journals The Effect of Slow-release Polymeric Derivatives of Auxins (NAA and 2,4-D) on Regeneration of Achimenes in Vitro

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 802D-802
Author(s):  
J.C. Vlahos ◽  
M. Dragassaki ◽  
A. Vasilaki ◽  
I. Assargiotaki ◽  
A.M. Tsatsakis
Keyword(s):  
Slow Release ◽  
Callus Formation ◽  
Dry Weight ◽  
Inhibitory Effects ◽  
Prolonged Action ◽  
Wide Range ◽  
Vigorous Growth ◽  
Derivatives Of ◽  
Different Levels

Polymeric formulations of plant growth regulators (PGR) are high-molecular weight systems in which the PGR unit can be slowly released providing prolonged action and effectiveness in a wide range of concentrations. In this study, Achimene explants were used for testing the biological activity of polymeric derivatives of NAA and 2,4-D. Shoots of Achimenes `Bella', obtained from leaf segments cultured in vitro, were transferred for 8 weeks on Murashige and Skoog (MS) medium supplemented with different levels of conventional or polymeric NAA (0, 0.01, 0.05, 0.1, 0.5, 1.0, 1.5, 2.0, 2.5 mg·liter–1) and 2.4-D (0, 0.01, 0.05, 0.1, 0.5, 1.0, 1.5 mg·liter–1), each combined with three levels of BAP (0, 0.1, 0.5 mg·liter–1). Compared to conventional NAA, twice as many shoots proliferated with higher dry weight, at 0.5 0.1, 1.0, or 1.5 mg·liter–1 polymeric NAA with 0.5 mg·liter–1 BAP. In another trial, the combination of 0.05 or 0.1 mg·liter–1 polymeric 2,4-D with 0.1 BAP gave more shoot and vigorous growth without callus formation, compared to conventional 2.4-D These results suggest that the polymeric derivatives of auxins used in this study enhance regeneration and growth of Achimenes in vitro more effectively than conventional formulations, at greater concentrations, without causing toxic or inhibitory effects.

scholarly journals Biodiversity and medicinal uses of globe artichoke (Cynara scolymus L.) plant

2019 ◽  
Vol 5 (1) ◽  
pp. 39-54 ◽  
Author(s):  
S Bekheet ◽  
V Sota
Keyword(s):  
Genetic Improvement ◽  
Genetic Erosion ◽  
Globe Artichoke ◽  
Crop Failure ◽  
Molecular Biology Technique ◽  
Medicinal Uses ◽  
Cynara Scolymus ◽  
Culture Techniques ◽  
Wide Range

Globe artichoke (Cynara scolymus L.) is a vegetable crop native of the Mediterranean basin and it is grown mainly for its immature flower buds. The edible parts of artichoke have important nutrition values to their content of inulin, fibers and minerals. Furthermore, artichoke is recognized as a medicinal plant where leaves and heads are rich source of polyphenolic compounds. It is recommended for the treatment of gallstones, liver disease or damage and poor liver function. The pharmaceutical properties of artichoke are linked to their special chemical composition, which includes high levels of polyphenols such as cynarin, along with its biosynthetic precursor chlorogenic acid. Globe artichoke is characterized by a wide range of biodiversity which represents a prerequisite for genetic improvement. However, there are many factors including biotic and abiotic stresses threatening the genetic resources of globe artichoke. Such factors could result in genetic erosion due to crop failure and loss of cultivars. Therefore, there is a great need to preserve the globe artichoke diversity for future genetic improvement. Because most cultivars are highly heterozygous, maintenance of globe artichoke germplasm in seed form is restricted. Otherwise, conservation in the field presents major drawbacks, which limit its efficacy and threaten the safety of germplasm conserved in this way. The application of biotechnology tools, i.e. plant tissue culture and molecular biology technique can provide reliable methods for supporting the exploitation and conservation of globe artichoke biodiversity. Furthermore, in vitro culture techniques could be used for the controlled production of artichoke secondary metabolites. This article discusses the biodiversity, conservation and nutritional and medicinal aspects of globe artichoke plant. J. Biodivers. Conserv. Bioresour. Manag. 2019, 5(1): 39-54

scholarly journals The Response of Callus Formation from Tacca Chantrieri Leaves with Various Concentrations of 2,4-D and BAP by In Vitro

2021 ◽  
Vol 9 (1) ◽  
pp. 8
Author(s):  
Maya Sari ◽  
Mayta Novaliza Isda
Keyword(s):  
Methanol Extract ◽  
Callus Formation ◽  
Southeast China ◽  
Culture Techniques ◽  
Tropical Plant ◽  
Inflorescence Morphology ◽  
The People ◽  
Randomized Design ◽  
Completely Randomized Design

The an annual herbaceous tropical plant which is one of the species of the genus Tacca from the Dioscoreaceae family is Tacca chantrieri. T. chantrieri has a unique inflorescence morphology like that of a bat. The people of Southeast China and Thailand have used by T. chantrieri rhizome as traditional medicine because the methanol extract contains secondary metabolites such as diarylheptanoids, pseudofurostan, withanolide, taccalonolide, and saponins. To maintain its sustainability, it is necessary to propagate T. chantrieri by using in vitro culture techniques such as callus culture. The purpose of this study was to determine the response of T. chantrieri leaf callus formation and to determine the optimal concentration with various concentrations of 2,4-D and BAP in vitro. This study used a completely randomized design consisting of control treatments, 1 and 2 mg L-1 2,4-D and concentrations of 0.5; 1.0; 1.5; 2.0 2,4-D combined with 3 mg L-1 BAP. The observations were made for 60 days after planting. The results showed that the concentration of 1.5 mg L-1 2,4-D + 3 mg L-1 BAP affected the percentage of live explants and the percentage of callus formation by 100% respectively, and the time of callus appeared 18.75 days after planting.

Darkening of Plant Tissues during in vitro Cultivation and Methods for its Prevention

2020 ◽  
Vol 36 (2) ◽  
pp. 26-42
Author(s):  
B.R. Kuluev
Keyword(s):  
Phenolic Compounds ◽  
Callus Formation ◽  
Plant Biotechnology ◽  
In Vitro Cultivation ◽  
Negative Effects ◽  
Wide Range ◽  
Negative Effect ◽  
Oxidative Transformations ◽  
Tissue Browning

One of the most common problems in the plant in vitro propagation is the tissue browning and subsequent necrosis, resulting from the oxidation of phenolic compounds, secondary metabolites produced in response to injury and released into the nutrient medium. This process is one of the main reasons for the decrease in the efficiency of callus formation, somatic embryogenesis, regeneration and genetic transformation of plants in vitro. Moreover, oxidative browning often leads to culture death. Therefore, the current problems in genetic and cellular engineering of a wide range of plant species can be solved only by preventing or reducing the negative effects of browning of in vitro cultures caused by the oxidative transformations of phenolic compounds into quinones toxic to cells. This review is devoted to the description of the main existing methods to prevent these adverse transformations. Various chemicals with antioxidant and adsorbing properties are used in plant biotechnology for this purpose, but there are no general approaches to solve the problem. Although the choice of the method to minimize the negative effect of phenolic compound oxidation depends, firs of all, on the species and variety of the plant, some agents, such as ascorbic acid, activated carbon, silver nitrate, can be considered as universal and quite effective in preventing oxidative darkening of explants in vitro. phenolic compounds, oxidative browning, polyphenol oxidase, tissue browning, in vitro, microclonal plant propagation The work was funded on the theme АААА-А17-117102740098-8.

scholarly journals Organogenic Nodule Formation in Hop: A Tool to Study Morphogenesis in Plants with Biotechnological and Medicinal Applications

2010 ◽  
Vol 2010 ◽  
pp. 1-16 ◽  
Author(s):  
Ana M. Fortes ◽  
Filipa Santos ◽  
Maria S. Pais
Keyword(s):  
Tissue Culture ◽  
Large Scale ◽  
Root Nodules ◽  
Molecular Networks ◽  
Nodule Formation ◽  
Culture Methods ◽  
Culture Techniques ◽  
Wide Range ◽  
Morphogenic Process

The usage ofHumulus lupulusfor brewing increased the demand for high-quality plant material. Simultaneously, hop has been used in traditional medicine and recently recognized with anticancer and anti-infective properties. Tissue culture techniques have been reported for a wide range of species, and open the prospect for propagation of disease-free, genetically uniform and massive amounts of plantsin vitro.Moreover, the development of large-scale culture methods using bioreactors enables the industrial production of secondary metabolites. Reliable and efficient tissue culture protocol for shoot regeneration through organogenic nodule formation was established for hop. The present review describes the histological, and biochemical changes occurring during this morphogenic process, together with an analysis of transcriptional and metabolic profiles. We also discuss the existence of common molecular factors among three different morphogenic processes: organogenic nodules and somatic embryogenesis, which strictly speaking depend exclusively on intrinsic developmental reprogramming, and legume nitrogen-fixing root nodules, which arises in response to symbiosis. The review of the key factors that participate in hop nodule organogenesis and the comparison with other morphogenic processes may have merit as a study presenting recent advances in complex molecular networks occurring during morphogenesis and together, these provide a rich framework for biotechnology applications.

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