In vitro scaling of Tagetes patula L. root culture

2020 ◽  
Vol 19 (1) ◽  
pp. 255-259
Author(s):  
L. P. Khlebova ◽  
E. S. Brovko
Keyword(s):  
Growth Rate ◽  
Hairy Roots ◽  
Growth Pattern ◽  
Nutrient Medium ◽  
Root Culture ◽  
Growth Cycle ◽  
Tagetes Patula ◽  
Laboratory Conditions

The features of the in vitro accumulation of biomass of Tagetes patula L. root culture in the process ofscaling under laboratory conditions in different volumes of the nutrient medium and culture vessels were studied. It wasrevealed that the growth pattern of the hairy roots line and its morphology depend on the volume of the incubation vessel.The pattern of increasing the growth rate and lengthening of the growth cycle of the culture in larger flasks (up to 500 ml)was established.

Kinetics of in vitro ageing of mouse embryo fibroblasts

1984 ◽  
Vol 65 (1) ◽  
pp. 163-175
Author(s):  
C. Karatza ◽  
W.D. Stein ◽  
S. Shall
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Mouse Embryo ◽  
Growth Pattern ◽  
Cell Strain ◽  
Growth Fraction ◽  
Cycle Duration ◽  
Cell Cycle Duration ◽  
Kinetics Of

The kinetics of ageing of normal mouse embryo fibroblast cells in culture have been determined. The growth pattern during every passage was established. It was observed that the growth pattern was not exponential, but that the growth rate declined progressively both within and with every passage. We also estimated the cell cycle parameters using the Fraction of Labelled Mitoses method at every passage. We found that the cell cycle duration was constant throughout the lifespan of this cell strain; the median value of the cell cycle duration was found to be 15.5 +/− 0.5 h (S.D., n = 8). From these two sets of observations we infer that the fraction of dividing cells declines smoothly from the beginning of the culture. Our data exclude quite positively any description of ageing of the fibroblast population in terms of a catastrophe or any abrupt change in the population. Our data are also inconsistent with a linear decline in growth fraction. On the contrary, we observed that there was a gradual and smooth decline in the growth rate of the strain, due to a smoothly declining growth fraction. This smooth change in the growth behaviour of this cell strain is accurately described by the mortalization theory of Shall & Stein in which the single parameter gamma (gamma), describes the change in reproductive potential over the entire lifespan. The parameter gamma describes the rate at which the doubling time of the culture increases. It is the number of generations at which half of the newborn cells are themselves reproductively sterile. Our present data provided an estimate of gamma for this cell strain, which was consistent during the entire lifespan of the strain; the best estimate of gamma for this cell strain was 20.3 +/− 0.6 generations (S.D., n = 19).

scholarly journals INFLUENCE OF NUTRITION COMPOSITIONS ON MICROCLONAL PROPAGATION DIFFERENT GENOTYPES OF THE WALNUT JUGLANS REGIA L.

2020 ◽  
Vol 1 (383) ◽  
pp. 105-112
Author(s):  
T. K. Yegizbayeva ◽  
T. V. Yausheva ◽  
S. N. Oleichenko ◽  
R. J. Licea-Moreno
Keyword(s):  
Growth Rate ◽  
Nutrient Medium ◽  
Juglans Regia ◽  
Growth Dynamics ◽  
Initial Stage ◽  
Positive Growth ◽  
Murashige Skoog ◽  
Mother Plants ◽  
Quality Material

Walnut is one of the most common nuts in many countries of the world. It is used in cooking and in traditional medicine. Walnut cultivation on an industrial scale in Kazakhstan began only in 2016-2017, so Kazakhstan scientists in this field do not yet have any scientific experience. The article presents the results of studies on the influence of the nutrient medium compositions Murashige-Skoog and Driver-Kunzhuki on the microclonal propagation of different genotypes of the walnut Juglans regia L. Two varieties were selected as mother plants: the Uzbek variety Ideal and the Chinese variety Liaohe-1, adapted in the southeast Kazakhstan. In the course of work, we compared the growth rate of walnut shoots of two varieties on nutrient media Murasige-Skoog (MS) and Driver-Kunzhuki (DKW) with a concentration of 6-benzylaminopurine (BAP) of 1 and 1.5 mg / l. After a month of cultivation of shoots, positive growth dynamics was observed only on MS medium. On plants of both varieties, the growth rate was slow on DKW medium, the plants had yellow leaves, some shoots blackened and perished. To obtain high-quality material, as well as to increase the growth rate of shoots, DKW medium was supplemented with FeEDDHA (119 mg / L) and phloroglucinol (50 mg / L). As a result of phenological observations: the height of the main shoot, the number of internodes, the formation of additional shoots, and the state of the plants, it was found that the best nutrient medium for microclonal propagation of walnut Ideal and Liaohe-1 is a modified DKW medium. MS can only be used at the initial stage - an introduction to in vitro culture.

scholarly journals Kultur akar rambut Cinchona ledgeriana dan C. succirubra dalam kultur in vitro Hairy root culture of Cinchona ledgeriana and C. succirubra by in vitro culture

2016 ◽  
Vol 72 (2) ◽  
Author(s):  
Nurita TORUAN-MATHIUS ◽  
. REFLINI ◽  
. NURHAIMI-HARIS ◽  
. JOKO-SANTOSO ◽  
A PRIANGANI-ROSWIEM
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Basal Medium ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Chain Reaction ◽  
Ri Plasmid ◽  
Polymerase Chain ◽  
Cinchona Ledgeriana

Summary Problems encountered in hairy root culture  of  C. ledgeriana and C. succirubra are low percentage of transformation of explants by Agrobacterium rhizogenes and slow growth of hairy root. The objective of this research was to evaluate the potential of several A. rhizogenes strains for initiation  hairy roots of  C. succirubra and C. ledgeriana, and to obtain the best medium for hairy root culture of Cinchona spesies. Axenic shoot and leaves explants of eight-month-old of C. ledgeriana and  C. succirubra seedlings were inoculated with A. rhizogenes strain ATCC-15834, ATCC-8196,    R-20001, 07-20001, A4, R-MAFFA, TISTR509, TISTR510 and LBA9457. Inoculated explants were cultured in solid MS medium with the addition of 100 mg/L amphicylin. Subculture of the hairy root was performed by transferred of root pieces into fresh liquid basal medium MS, B5, White and Heller. Hairy roots from the best of basal medium were subcultured on the same medium with the addition of 50  and 100 mg/L   L-tryptophane, three or five times concentration of MS vitamins. The integration of T-DNA of   A. rhizogenes in hairy root was confirmed with specific primer for TL and TR-DNA of plasmid by Polymerase Chain Reaction analysis. The results showed that only A. rhizogenes strain  LBA 9457 were effective for  transformation of explants from both Cinchona species. The fastest hairy roots growth were found  in MS medium, while growth in others medium was poor. Hairy roots of  C. ledgeriana has vigor and growth better than hairy roots of C. succirubra. MS with the addition of 50 mg/L  L-tryptophane and  three times the concen-trations of vitamin  is the best medium for hairy root growth and vigor. Hairy roots of  C. succirubra and C. ledgeriana used in this studies were confirmed that hairy roots  contained TL and TR-DNA region of Ri plasmid with molecular weight 780 and 1600 bp.  The results showed that strain of A. rhizogenes, plant species, source of explant and composition of medium affect the initiation, growth, development  and vigor of hairy roots.Ringkasan Masalah dalam kultur akar rambut  C. ledgeriana dan C. succirubra adalah rendahnya tingkat keberhasilan transformasi eksplan dengan Agrobacterium rhizogenesdan pertumbuhannya yang lambat. Penelitian ini bertujuan untuk mengevaluasi  potensi dari beberapa galur A. Rhizogenes untuk inisiasi, mendapatkan komposisi medium terbaik untuk pertumbuhan akar rambut C. ledgeriana dan C. succirubra, serta konfirmasi terintegrasinya TR dan TL-DNA Ri plasmid ke dalam jaringan eksplan.  Eksplan batang  dan  daun  berasal  dari kecambah aksenik C. ledgeriana dan C. succirubra berumur delapan bulan diinokulasi dengan A. rhizogenes galur 15834, 8196, R-20001, 07-20001, A4, R.MAFFA,TISTR 509, TISTR 510 dan LBA 9457. Eksplan yang sudah diinokulasi dikulturkan dalam medium MS padat. Subkultur dilakukan dengan cara mentransfer potongan ujung akar rambut ke dalam medium cair MS, B5, White dan Heller. Akar rambut dari medium kultur yang terbaik kemudian disubkultur ke dalam medium yang sama dengan penambahan 50 dan 100 mg/L L-triptofan dengan konsentrasi vitamin sebanyak tiga kali dan lima kali dari konsentrasi normal MS. Integrasi T-DNA dalam akar rambut dikonfirmasi meng-gunakan Polymerase Chain Reaction  dengan primer spesifik untuk TL dan TR-DNA plasmid. Hasil yang diperoleh menunjukkan bahwa hanya A.rhizogenes galur LB9457 yang efektif menginfeksi eksplan baik batang maupun daun dari kedua spesies kina. Induksi, pertumbuhan dan vigor akar rambut yang terbaik diperoleh dari medium MS dengan penambahan 50 mg/L L-triptofan dan tiga kali konsentrasi vitamin. Hasil konfirmasi akar rambut baik dari batang maupun daun menggunakan PCR, menunjukkan bahwa TL dan TR-DNA dari Ri plasmid  A. rhizogenes mampu menghasilkan pita-pita DNA dengan BM780 dan 1600 pb. Hasil yang diperoleh menunjukkan bahwa galur  A. rhizogenes, spesies tanaman, sumber eksplan dan komposisi medium berpengaruh terhadap inisiasi, pertumbuhan,  perkembangan dan vigor akar rambut.

scholarly journals Induction of transgenic hairy roots in soybean genotypes by Agrobacterium rhizogenes‑mediated transformation

2011 ◽  
Vol 46 (9) ◽  
pp. 1070-1075 ◽  
Author(s):  
Ricardo Luís Mayer Weber ◽  
Maria Helena Bodanese‑Zanettini
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Fluorescent Protein ◽  
Root Culture ◽  
Transformation Frequency ◽  
Plant Production ◽  
Uniform Size ◽  
Soybean Genotypes

The objective of this work was to perform the screening of soybean genotypes as to their ability to respond to the induction of hairy roots by Agrobacterium rhizogenes‑mediated transformation. Four Brazilian soybean cultivars (BRSMG 68 Vencedora, BRS 137, Embrapa 48, and MG/BR 46 Conquista) and two North American ones adapted to Brazilian cropping conditions (Bragg and IAS‑5) were screened for their capacity to respond to A. rhizogenes in protocols for in vitro hairy root culture and ex vitro composite plant production. Four‑day‑old seedlings with uniform size were injected with A. rhizogenes harboring the plasmid p35S‑GFP. Seedlings expressing green fluorescent protein (GFP) in at least one hairy root were used to determine the transformation frequency. Using an axenic in vitro protocol, excised cotyledons from four‑day‑old seedlings were infected with A. rhizogenes harboring the pCAMBIA1301 plasmid, containing the gusA reporter gene. The transformation frequency and the number of days for hairy root emergence after bacterial infection (DAI) were evaluated. The transformation frequency and DAI varied according to the genotype. Cultivars MG/BR 46 Conquista and BRSMG 68 Vencedora are more susceptible to A. rhizogenes and can be recommended for transformation experiments.

Physiological Effects of Sucrose Substitutes and Artificial Sweeteners on Growth Pattern and Acid Production of Glucose-Grown Streptococcus mutans Strains in vitro

1976 ◽  
Vol 31 (5-6) ◽  
pp. 245-251 ◽  
Author(s):  
Harald A. B. Linke ◽  
Catherine A. Chang
Keyword(s):  
Growth Rate ◽  
Streptococcus Mutans ◽  
Growth Pattern ◽  
Acid Production ◽  
High Rate ◽  
Artificial Sweeteners ◽  
Glucose Fermentation ◽  
Rate Of Growth ◽  
Moderate Growth

Abstract The synergistic effects of four sucrose substitutes, polysorbate and five artificial sweeteners were studied in vitro on growth pattern and acid production of seven glucose-grown Streptococcus mutans strains, representing the five serological groups after Bratthall. Four distinct growth pat­ terns during glucose fermentation were observed: high rate of growth with low acid production, moderate growth rate with moderate acid production, moderate growth rate with high acid pro­duction, and slow rate of growth with moderate acid production. Depending on the strain used, the final OD at 546 nm ranged from 0.55 to 0.99 and the final pH of the medium varied between 4.65 and 4.15. While added sucrose substitutes, with exceptions, usually enhanced growth rate, most artificial sweeteners suppressed or, at higher concentrations, even inhibited growth of S. mutans-, addition of polysorbate to the medium always increased growth rate of S. mutans significantly. The presence of sucrose substitutes during glucose fermentation had no effect on final pH of the medium, but addition of artificial sweeteners, especially sodium saccharin, elevated final pH up to 1.8 units. The observed physiological patterns and differences within the several strains of S. mutans during glucose fermentation in vitro do not necessarily relate to the five serological groups of the species.

scholarly journals TÉCNICAS DE TRANSFORMAÇÃO E LINHAGENS De Agrobacterium rhizogenes NA PRODUÇÃO DE HAIRY ROOTS DE Ocimum basilicum L.

Nativa ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 266
Author(s):  
José Nicomedes Júnior ◽  
Roberta Cristiane Ribeiro ◽  
Luis Louro Berbara ◽  
Elvia Mariam Lis Martinez Stark ◽  
Wagner Campos Otoni ◽  
...  
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Leaf Explants ◽  
Laboratory Strain ◽  
Root Culture ◽  
Ocimum Basilicum ◽  
Best Response ◽  
Agrobacterium Rhizogenes Strains ◽  
Whole Plants

O manjericão é uma planta medicinal e seus metabólitos especiais são utilizados com fins terapêuticos e industriais. Este trabalho teve por objetivo avaliar técnicas e linhagens de Agrobacterium rhizogenes na produção de raízes transformadas de duas variedades de Ocimum basilicum L., Dark Opal (roxo) e Minete Anão (verde). Verificou-se o efeito da inoculação com linhagens de A. rhizogenes selvagens (A4, 8196, 9402, 2659, 2659 G, 17242, LBA, 15834) ou transformadas (R1601) em plantas inteiras ou explantes (folhas e segmentos de caule) de manjericão, cultivados em casa de vegetação ou in vitro. A inoculação com as linhagens LBA e 8196 possibilitou redução no número de explantes de folha de manjericão-verde oxidados, quando comparado ao controle. Dentre as linhagens selvagens, as que proporcionaram melhor indução na produção de raízes transformadas foram A4, LBA e 8196, respectivamente. Entretanto, a cepa R1601 apresentou a melhor resposta. Raízes foram eficientemente induzidas mediante a inoculação de explantes de manjericão-roxo e manjericão-verde e expressaram o fenótipo típico de raízes em cabeleira (hairy roots). As culturas clonais de manjericão-verde apresentaram rápido crescimento em meio de cultura livre de reguladores de crescimento.Palavras-chave: agrobactéria, cultura de raízes, manjericão, plantas medicinais. TRANSFORMATION TECHNIQUES AND Agrobacterium rhizogenes Strains In The Production Of Hairy Roots Of Ocimum basilicum L. ABSTRACT:Basil is a medicinal plant and its special metabolites are used for therapeutic or industrial purposes. The aim of this work was to evaluate Agrobacterium rhizogenes techniques and strains in the production of hairy roots of two varieties of Ocimum basilicum L., Dark Opal (purple) and Dwarf Racer (green). The effects of inoculation of A. rhizogene (A4, 8196, 9402, 2659, 2659, 2659 G, 17242, LBA, 15834) or (R1601) lines on whole plants or explants (leaves and stem segments) of basil, grown in a greenhouse or in vitro, in the production of hairy roots and calluses made by subcultures. Inoculation with the LBA and 8196 lines allowed a reduction in the number of oxidized basil-green leaf explants when compared to the control. Among the wild strains, those that had best ability to induce hairy root of hairy roots were A4, LBA and 8196, respectively. However, the R1601 laboratory strain had the best response. Roots were efficiently induced by the inoculation of explants basil-purple and basil-green and expressed the typical phenotype of hairy roots. Green-basal clonal cultures showed rapid growth in culture medium free of growth regulators.Keywords: agrobacterim, basil, medicinal plants, root culture. DOI:

scholarly journals Kultur akar rambut Cinchona ledgeriana dan C. succirubra dalam kultur in vitro Hairy root culture of Cinchona ledgeriana and C. succirubra by in vitro culture

2016 ◽  
Vol 72 (2) ◽  
Author(s):  
Nurita TORUAN-MATHIUS ◽  
. REFLINI ◽  
. NURHAIMI-HARIS ◽  
. JOKO-SANTOSO ◽  
A PRIANGANI-ROSWIEM
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Basal Medium ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Chain Reaction ◽  
Ri Plasmid ◽  
Polymerase Chain ◽  
Cinchona Ledgeriana

Summary Problems encountered in hairy root culture  of  C. ledgeriana and C. succirubra are low percentage of transformation of explants by Agrobacterium rhizogenes and slow growth of hairy root. The objective of this research was to evaluate the potential of several A. rhizogenes strains for initiation  hairy roots of  C. succirubra and C. ledgeriana, and to obtain the best medium for hairy root culture of Cinchona spesies. Axenic shoot and leaves explants of eight-month-old of C. ledgeriana and  C. succirubra seedlings were inoculated with A. rhizogenes strain ATCC-15834, ATCC-8196,    R-20001, 07-20001, A4, R-MAFFA, TISTR509, TISTR510 and LBA9457. Inoculated explants were cultured in solid MS medium with the addition of 100 mg/L amphicylin. Subculture of the hairy root was performed by transferred of root pieces into fresh liquid basal medium MS, B5, White and Heller. Hairy roots from the best of basal medium were subcultured on the same medium with the addition of 50  and 100 mg/L   L-tryptophane, three or five times concentration of MS vitamins. The integration of T-DNA of   A. rhizogenes in hairy root was confirmed with specific primer for TL and TR-DNA of plasmid by Polymerase Chain Reaction analysis. The results showed that only A. rhizogenes strain  LBA 9457 were effective for  transformation of explants from both Cinchona species. The fastest hairy roots growth were found  in MS medium, while growth in others medium was poor. Hairy roots of  C. ledgeriana has vigor and growth better than hairy roots of C. succirubra. MS with the addition of 50 mg/L  L-tryptophane and  three times the concen-trations of vitamin  is the best medium for hairy root growth and vigor. Hairy roots of  C. succirubra and C. ledgeriana used in this studies were confirmed that hairy roots  contained TL and TR-DNA region of Ri plasmid with molecular weight 780 and 1600 bp.  The results showed that strain of A. rhizogenes, plant species, source of explant and composition of medium affect the initiation, growth, development  and vigor of hairy roots.Ringkasan Masalah dalam kultur akar rambut  C. ledgeriana dan C. succirubra adalah rendahnya tingkat keberhasilan transformasi eksplan dengan Agrobacterium rhizogenesdan pertumbuhannya yang lambat. Penelitian ini bertujuan untuk mengevaluasi  potensi dari beberapa galur A. Rhizogenes untuk inisiasi, mendapatkan komposisi medium terbaik untuk pertumbuhan akar rambut C. ledgeriana dan C. succirubra, serta konfirmasi terintegrasinya TR dan TL-DNA Ri plasmid ke dalam jaringan eksplan.  Eksplan batang  dan  daun  berasal  dari kecambah aksenik C. ledgeriana dan C. succirubra berumur delapan bulan diinokulasi dengan A. rhizogenes galur 15834, 8196, R-20001, 07-20001, A4, R.MAFFA,TISTR 509, TISTR 510 dan LBA 9457. Eksplan yang sudah diinokulasi dikulturkan dalam medium MS padat. Subkultur dilakukan dengan cara mentransfer potongan ujung akar rambut ke dalam medium cair MS, B5, White dan Heller. Akar rambut dari medium kultur yang terbaik kemudian disubkultur ke dalam medium yang sama dengan penambahan 50 dan 100 mg/L L-triptofan dengan konsentrasi vitamin sebanyak tiga kali dan lima kali dari konsentrasi normal MS. Integrasi T-DNA dalam akar rambut dikonfirmasi meng-gunakan Polymerase Chain Reaction  dengan primer spesifik untuk TL dan TR-DNA plasmid. Hasil yang diperoleh menunjukkan bahwa hanya A.rhizogenes galur LB9457 yang efektif menginfeksi eksplan baik batang maupun daun dari kedua spesies kina. Induksi, pertumbuhan dan vigor akar rambut yang terbaik diperoleh dari medium MS dengan penambahan 50 mg/L L-triptofan dan tiga kali konsentrasi vitamin. Hasil konfirmasi akar rambut baik dari batang maupun daun menggunakan PCR, menunjukkan bahwa TL dan TR-DNA dari Ri plasmid  A. rhizogenes mampu menghasilkan pita-pita DNA dengan BM780 dan 1600 pb. Hasil yang diperoleh menunjukkan bahwa galur  A. rhizogenes, spesies tanaman, sumber eksplan dan komposisi medium berpengaruh terhadap inisiasi, pertumbuhan,  perkembangan dan vigor akar rambut.

scholarly journals Characteristics growth of mycelium of mushroom shiitake Lentinula Edodes (Berk.) Pegler in vitro

2019 ◽  
pp. 20-25
Author(s):  
T. Ivanova ◽  
N. Voloschuk
Keyword(s):  
Growth Rate ◽  
Light Microscopy ◽  
Pure Culture ◽  
Nutrient Medium ◽  
Sodium Selenite ◽  
Lentinula Edodes ◽  
Ph Value ◽  
Nutrient Media ◽  
Microbiological Methods

Goal. The purpose of this work is to study the growth of the mycelium of the fungus Lentinula Edodes (Berk.) Pegler on nutrient media of different composition and also to study the features of the use of Avatar-1 micro fertilizer and sodium selenite. Methods. Biotechnological research methods. Sodium selenite (Na2S eO3) at a concentration of 1.0 mmol / l was also used. Pure culture of the mushrooms had a very dense structure and white color. In our work, we used biotechnological methods — obtain this by subculturing the L. edodes strain in vitro; microbiological methods — obtaining pure culture of the fungus, the study of the cultural properties of the colonies. We set the pH value (pH) of nutrient media at the beginning and at the end of incubation. We applied mycological methods to measure the speed, density of growth and dry mass of mycelium. we used the light microscopy method. We performed statistical data processing.Method of light microscopy. Results. The experiments showed about acceleration of mycelial growth, mass and the greatest yield of mycelium L. edodes were on a nutrient medium with microfertilizer Avatar-1. In the experiment, it was found that the maximum overgrowth of the medium by mycelium occurs at 7 days. We have been proved that in the «Avatar-1» nutrient medium there was an increase and consolidation of bifurcated hyphae and buckles of L. edodes. There was even germination of mycelium that did not have too thin or thick hyphae. The dependence of growth rate on the type of nutrient medium. Also, the of doses of the drug, which effectively influences the technology of obtaining primary mycelium L. edodes. Conclusions. The dependence of the growth rate on the type of nutrient medium, the dose administration rate, as well as the cultivation regimes, that effectively influence and cheapen the technology of obtaining the primary mycelium L. edodes is demonstrated.

Assessing the growth and saponin accumulation in hairy and adventitious roots of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.)

2016 ◽  
Vol 14 (2) ◽  
pp. 231-236
Author(s):  
Trịnh Thị Hương ◽  
Phạm Bích Ngọc ◽  
Chu Hoàng Hà ◽  
Dương Tấn Nhựt
Keyword(s):  
Growth Rate ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Hairy Roots ◽  
Growth Regulators ◽  
Adventitious Roots ◽  
Dry Matter ◽  
Ginseng Root ◽  
Free Medium

In this study, adventitious and hairy roots of Vietnamese ginseng were used to assess the ability of growth and saponin accumulation. Adventitious roots were derived from leaf samples in vitro (1.0 x 1.0 cm of size) cultured on SH medium supplemented with 5.0 mg.l-1 IBA, 30 g.l-1 sucrose, 8 g.l-1 agar, pH 5.8, and subcultured on the same medium for multiplication. Hairy roots were derived from callus infected with Agrobacterium rhizogenes strain ATCC 15834, then these roots were cultured on plant growth regulators-free SH medium supplemented with 50 g.l-1 sucrose, 8 g.l-1 agar, pH 5.8. During the early culture of two months, the results showed that the growth rate of hairy roots was lower than that of adventitious roots. However, in the later period of culture, the growth rate of hairy roots was higher than that of adventitious roots. After 5 months of culture, the growth rate of hairy roots was 20.87 times and they kept growing as well as branching, while the growth rate of adventitious roots was only 13.52 times and they did not grow further after three months of culture. Analytical results showed that the total saponins of total dry matter of hairy roots (0.101 mg) were higher than that of adventitious roots (0.0681 mg). The main ginsenoside of hairy roots (MR2) was also higher than that of adventitious roots 3.03 fold. In addition, the hairy roots grew on plant growth regulators-free medium while adventitious roots grew on medium supplemented with auxin. Therefore, hairy roots proved to be suitable source material for Vietnamese ginseng root biomass production in the bioreactor systems.

scholarly journals Analysis of individual biologically active compounds from root crop extracts of purple alfalfa (medicago sativa) in vitro

2021 ◽  
Vol 7 (2) ◽  
pp. 564-574
Author(s):  
Irina Sergeevna Milentyeva ◽  
Anastasiya Igorevna Dmitrieva ◽  
Lyudmila Konstantinovna Asyakina ◽  
Yulia Vladimirovna Golubtsova
Keyword(s):  
Medicago Sativa ◽  
Growth Index ◽  
Root Culture ◽  
Growth Cycle ◽  
Biologically Active ◽  
Root Cultures ◽  
Degree Of Extraction ◽  
Degree Of Purification ◽  
Active Substances

The authors selected the parameters for obtaining root cultures in vitro Medicago sativa. The number of cell culture lines studied is 15. The duration of the growth cycle is less than 50 days. The number of processed explants is 75. The growth characteristics of root cultures of in vitro purple alfalfa (Medicago sativa) were studied. The growth index for the dry biomass of the in vitro root culture of alfalfa was 31. A qualitative and quantitative analysis of the composition of BAS in the biomass of the obtained root cultures of in vitro alfalfa (Medicago sativa) was carried out. Unique biologically active substances (not previously described for alfalfa) were identified.): apigenin, naringenin, myricetin, chrysoeriol, coumestrol, and scopoletin. Rational parameters of isolation of individual BAS from the extract of root cultures of in vitro alfalfa (Medicago sativa) were selected. The degree of extraction of individual BAS is 80%. Methods of purification of individual BAS Medicago sativa have been developed. The degree of purification of individual BAS is 95%.

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