Rapid plant regeneration, analysis of genetic fidelity, and neoandrographolide content of micropropagated plants of Andrographis alata (Vahl) Nees

Abstract Background Andrographis alata (Vahl) Nees is a medicinal plant which was reported to have the highest concentration of neoandrographolide that has several therapeutic values. Natural populations of Andrographis alata are dwindling due to destruction of natural habitat and over exploitation. Therefore, in vitro propagation of Andrographis alata was undertaken, and successful method is presented here. Results Micropropagation of Andrographis alata was realized on MS nutrient medium augmented with BAP (10 μM), and multiple shoots were regenerated from nodal explants. Induction of roots was attained from shoots on ¼ concentration of MS nutrient medium supplemented with IBA (1 μM). Randomly amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analysis showed that there is genetic fidelity in the regenerated plants. Reverse phase high performance liquid chromatographic analysis of regenerated plants showed the presence of neoandrographolide, equivalent to that of mother plants. Conclusions Successful in vitro regeneration of Andrographis alata is presented here, and it is quite useful for its mass multiplication. The micropropagated plants are useful for restoration of plants in nature and for utilization by the pharmaceutical industry for extraction of neoandrographolide.

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In Vitro Propagation of an Endangered Helianthus verticillatus by Axillary Bud Proliferation

Plants ◽

10.3390/plants9060712 ◽

2020 ◽

Vol 9 (6) ◽

pp. 712

Author(s):

Marzena Nowakowska ◽

Žaklina Pavlović ◽

Marcin Nowicki ◽

Sarah L. Boggess ◽

Robert N. Trigiano

Keyword(s):

Endangered Species ◽

Shoot Proliferation ◽

Genetic Fidelity ◽

Induction Medium ◽

Axillary Shoot ◽

Axillary Shoot Proliferation ◽

Regenerated Plants ◽

Micropropagated Plants ◽

Ssrs Markers

Helianthus verticillatus (Asteraceae), whorled sunflower, is a perennial species restricted to a few locations in the Southeastern United States. Habitat loss has caused H. verticillatus to become rare, and since 2014, it has been federally listed as an endangered species. As a part of the recovery plan for the restoration and protection of H. verticillatus, an efficient micropropagation protocol based on axillary shoot proliferation was developed. Various concentrations of 6-benzylaminopurine (BAP; 0 to 4.44 µM) were examined for their morphogenetic potential in the regeneration of six genotypes of H. verticillatus from the nodal explants derived from greenhouse-grown plants. Both the BAP concentration and genotype had significant effects on the regeneration capacity of H. verticillatus. Although the induced buds were observed on ½-strength Murashige and Skoog medium without plant growth regulators, a higher rate of induction and bud development were achieved on media with either 0.88 or 2.22 µM BAP, regardless of the genotype. Successful rooting of the induced shoots was achieved within four weeks after the transfer from the induction medium to the fresh ½-strength MS medium, but the rooting efficiency was dependent on the plant’s genetic background. Regenerated plantlets, with well-developed shoots and roots, were acclimatized successfully to greenhouse conditions with a 97% survival rate. Simple sequence repeats (SSRs) markers were employed to assess the genetic uniformity of the micropropagated plants of H. verticillatus. No extraneous bands were detected between regenerants and their respective donor plants, confirming the genetic fidelity and stability of regenerated plants. To our knowledge, the protocol developed in this study is the first such report for this endangered species.

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In Vitro Regeneration of Phyllanthus amarus Schum. and Thonn.: An Important Medicinal Plant

Our Nature ◽

10.3126/on.v7i1.2557 ◽

1970 ◽

Vol 7 (1) ◽

pp. 110-115 ◽

Cited By ~ 3

Author(s):

A. Sen ◽

M.M. Sharma ◽

D. Grover ◽

A. Batra

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Shoot Elongation ◽

Nodal Segments ◽

Nodal Segment ◽

Phyllanthus Amarus ◽

Regenerated Plants ◽

Half Strength ◽

Important Medicinal Plant

An efficient in vitro plant regeneration protocol was developed for the medicinally potent plant species Phyllanthus amarus Schum. and Thonn. (Euphorbiaceae) using nodal segment as explant. Maximum multiplication of shoots (15.275±0.96) was achieved on Murashige and Skoog’s medium supplemented with BAP (0.5 mg/l) after 3-4 weeks of inoculation. The shoots were separated from cluster and subcultured for their elongation on the same medium. In vitro flowering was also observed on the elongated shoots after 3–4 weeks of sub culturing on the shoot elongation medium. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l). Regenerated plants were successfully hardened and acclimatized, 80 % of plantlets survived well under natural conditions after transplantation.Key words: In vitro regeneration, multiple shoots, nodal segments, Phyllanthus amarusDOI: 10.3126/on.v7i1.2557Our Nature (2009) 7:110-115

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In vitro regeneration, antioxidant potential, and genetic fidelity analysis of Asystasia gangetica (L.) T.Anderson

In Vitro Cellular & Developmental Biology - Plant ◽

10.1007/s11627-020-10141-5 ◽

2021 ◽

Author(s):

Abhirami Dilkalal ◽

Annapurna A S ◽

Umesh T G

Keyword(s):

In Vitro Regeneration ◽

Genetic Fidelity ◽

Antioxidant Potential ◽

Asystasia Gangetica

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Metabolic profiling, in vitro propagation, and genetic assessment of the endangered rare plant Anarrhinum pubescens

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00210-6 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Asmaa Abdelsalam ◽

Ehab Mahran ◽

Kamal Chowdhury ◽

Arezue Boroujerdi

Keyword(s):

In Vitro Propagation ◽

Genetic Fidelity ◽

Ex Situ ◽

Wild Plant ◽

Rare Plant ◽

Chemical Profiling ◽

Nodal Cutting ◽

Regenerated Plants ◽

Plant Shoots

Abstract Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.

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Cytokinin Mediated Increased In Vitro Production of Secondary Metabolites with Special Reference to Solasodine in Solanum erianthum

Planta Medica International Open ◽

10.1055/a-1484-9750 ◽

2021 ◽

Vol 8 (02) ◽

pp. e62-e68

Author(s):

Jeeta Sarkar ◽

Nirmalya Banerjee

Keyword(s):

Secondary Metabolites ◽

High Performance ◽

Culture Medium ◽

In Vitro Production ◽

Leaf Extracts ◽

Plant Parts ◽

Regenerated Plants ◽

Vitro Production ◽

Regenerated Plantlets

AbstractSteroid alkaloid solasodine is a nitrogen analogue of diosgenin and has great importance in the production of steroidal medicines. Solanum erianthum D. Don (Solanaceae) is a good source of solasodine. The aim of this study was to evaluate the effect of different cytokinins on the production of secondary metabolites, especially solasodine in the in vitro culture of S. erianthum. For solasodine estimation, field-grown plant parts and in vitro tissues were extracted thrice and subjected to high-performance liquid Chromatography. Quantitative analysis of different secondary metabolites showed that the amount was higher in the in vitro regenerated plantlets compared to callus and field-grown plants. The present study critically evaluates the effect of the type of cytokinin used in the culture medium on solasodine accumulation in regenerated plants. The highest solasodine content (46.78±3.23 mg g-1) was recorded in leaf extracts of the in vitro grown plantlets in the presence of 6-γ,γ-dimethylallylamino purine in the culture medium and the content was 3.8-fold higher compared to the mother plant.

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In vitro regeneration of Persian poppy (Papaver bracteatum)

Biologia ◽

10.2478/s11756-010-0079-6 ◽

2010 ◽

Vol 65 (4) ◽

Cited By ~ 4

Author(s):

Sara Rostampour ◽

Haleh Sohi ◽

Ali Dehestani

Keyword(s):

Ascorbic Acid ◽

Callus Induction ◽

High Performance ◽

High Efficiency ◽

In Vitro Regeneration ◽

Naphthalene Acetic Acid ◽

Dichlorophenoxyacetic Acid ◽

Papaver Bracteatum ◽

Callus Proliferation

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.

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ASSESSMENT OF GENETIC STABILITY OF MICROPROPAGATED Eucalyptus globulus Labill HYBRID CLONES BY MEANS OF FLOW CYTOMETRY AND MICROSATELLITES MARKERS

Revista Árvore ◽

10.1590/1806-90882017000100014 ◽

2017 ◽

Vol 41 (1) ◽

Author(s):

Leandro Silva Oliveira ◽

Aloisio Xavier ◽

Wagner Campos Otoni ◽

José Marcello Salabert Campos ◽

Lyderson Facio Viccini ◽

...

Keyword(s):

Flow Cytometry ◽

Microsatellite Markers ◽

Genetic Stability ◽

Culture Medium ◽

Eucalyptus Grandis ◽

Genetic Fidelity ◽

Shoot Multiplication ◽

Micropropagated Plants ◽

Hybrid Clones

ABSTRACT Flow cytometry and microsatellite markers were used to determine a genetic fidelity of micropropagated plants from the two Eucalyptus urophylla x E. globulus clones and a Eucalyptus grandis x E. globulus clone derived from adult material. Clones were repeatedly subcultured for 25 subcultures on MS medium supplemented with BA (2.22 µM) and ANA (0.05 µM) for in vitro shoot multiplication. The elongation was performed in MS culture medium supplemented with AIB (2.46 µM) and BA(0.22 µM). The ex vitro rooting and acclimatization phases were lead at the same time. The micropropagated clones showed genetic stability by flow cytometry and microsatellite markers. The results proved that micropropagation, for purposes of rejuvenation, can be a viable technique to generate genetically stable or identical E. globulus hybrid clones.

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