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

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.

scholarly journals In Vitro Propagation of Rhododendron tomentosum – an Endangered Essential Oil Bearing Plant from Peatland

2016 ◽  
Vol 58 (2) ◽  
pp. 29-43 ◽  
Author(s):  
Anna Jesionek ◽  
Adam Kokotkiewicz ◽  
Paulina Wlodarska ◽  
Natalia Filipowicz ◽  
Adam Bogdan ◽  
...  
Keyword(s):  
Essential Oil ◽  
Random Amplified Polymorphic Dna ◽  
Continuous Production ◽  
Leaf Explants ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Regenerated Plants ◽  
Rhododendron Tomentosum

Abstract Rhododendron tomentosum Harmaja (formerly Ledum palustre L.) is a medicinal peat bog plant native to northern Europe, Asia and North America. This plant has a distinctive aroma thanks to the presence of essential oil, to which it also owes its anti-inflammatory, analgesic, antimicrobial and insecticidal properties. However, in Europe R. tomentosum is classified as an endangered species, mainly due to degradation of peatlands. In the present work, the micropropagation protocol for R. tomentosum was established for the first time, providing both an ex situ conservation tool and a means of continuous production of in vivo and in vitro plant material for further studies. R. tomentosum microshoots were initiated from leaf explants and further multiplied using Schenk-Hildebrandt (SH) medium supplemented with 9.84 μM 2iP and 1.00 μM TDZ. The shoots were elongated on the SH medium supplemented with 24.6 μM 2iP and subsequently rooted using the perlite substrate saturated with half-strength Woody Plant medium supplemented with 1.0% sucrose and 4.92 μM IBA. The regenerated plants were hardened on the phytohormone-free SH medium and acclimatized using 3:1:1 deacidified peat:perlite:gravel substrate. The identity of the mother plant was confirmed at morphological and molecular levels and Random Amplified Polymorphic DNA (RAPD) method was implemented to assess the genetic fidelity of the regenerants. The essential oil content of the maternal plant, in vitro shoots and the regenerants was determined by steam-distillation, and the obtained volatile fractions were analyzed by GC/MS.

scholarly journals Genetic Fidelity Study of the in vitro Regenerated Plants in LA Hybrids of Lilium cv. Pavia

2018 ◽  
Vol 7 (09) ◽  
pp. 1223-1231 ◽  
Author(s):  
Asm ita ◽  
S.S. Sindhu ◽  
M. Jayanthi ◽  
M.R. Dhiman ◽  
M.K. Singh ◽  
...  
Keyword(s):  
Genetic Fidelity ◽  
Regenerated Plants

In vitro propagation and genetic fidelity analysis of alginate-encapsulated Bacopa monnieri shoot tips using Gracilaria salicornia extracts

2016 ◽  
Vol 29 (1) ◽  
pp. 481-494 ◽  
Author(s):  
Arockiam Sagina Rency ◽  
Lakkakula Satish ◽  
Subramani Pandian ◽  
Periyasamy Rathinapriya ◽  
Manikandan Ramesh
Keyword(s):  
In Vitro Propagation ◽  
Genetic Fidelity ◽  
Bacopa Monnieri ◽  
Shoot Tips ◽  
Gracilaria Salicornia

scholarly journals Application of the MSAP Technique to Evaluate Epigenetic Changes in Plant Conservation

2020 ◽  
Vol 21 (20) ◽  
pp. 7459
Author(s):  
María Elena González-Benito ◽  
Miguel Ángel Ibáñez ◽  
Michela Pirredda ◽  
Sara Mira ◽  
Carmen Martín
Keyword(s):  
Dna Methylation ◽  
In Situ Conservation ◽  
Plant Conservation ◽  
Ex Situ ◽  
Conservation Strategies ◽  
Regenerated Plants ◽  
Before And After ◽  
Methylation Patterns

Epigenetic variation, and particularly DNA methylation, is involved in plasticity and responses to changes in the environment. Conservation biology studies have focused on the measurement of this variation to establish demographic parameters, diversity levels and population structure to design the appropriate conservation strategies. However, in ex situ conservation approaches, the main objective is to guarantee the characteristics of the conserved material (phenotype and epi-genetic). We review the use of the Methylation Sensitive Amplified Polymorphism (MSAP) technique to detect changes in the DNA methylation patterns of plant material conserved by the main ex situ plant conservation methods: seed banks, in vitro slow growth and cryopreservation. Comparison of DNA methylation patterns before and after conservation is a useful tool to check the fidelity of the regenerated plants, and, at the same time, may be related with other genetic variations that might appear during the conservation process (i.e., somaclonal variation). Analyses of MSAP profiles can be useful in the management of ex situ plant conservation but differs in the approach used in the in situ conservation. Likewise, an easy-to-use methodology is necessary for a rapid interpretation of data, in order to be readily implemented by conservation managers.

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

Plants ◽  
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.

GIS-facilitated in vitro propagation and ex situ conservation of Achillea occulta

2011 ◽  
Vol 107 (3) ◽  
pp. 531-540 ◽  
Author(s):  
Katerina Grigoriadou ◽  
Nikos Krigas ◽  
Eleni Maloupa
Keyword(s):  
In Vitro Propagation ◽  
Ex Situ Conservation ◽  
Ex Situ

scholarly journals Nutrient optimization for improved in vitro plant regeneration in Eclipta alba (L.) Hassk. and assessment of genetic fidelity using RAPD analysis

2015 ◽  
Vol 24 (2) ◽  
pp. 223-234
Author(s):  
Shruti Bardar ◽  
Varsha Khurana Kaul ◽  
Sumita Kachhwaha ◽  
SL Kothari
Keyword(s):  
Basal Medium ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Nutrient Level ◽  
Induction Medium ◽  
Eclipta Alba ◽  
Shoot Bud ◽  
Bud Induction ◽  
Regeneration Response

This study highlights the effect of different inorganic micronutrients like copper, cobalt, molybdenum, zinc, boron, iodine, iron and manganese in accelerating and amplifying in vitro shoot bud induction and proliferation of a medicinally important plant, Eclipta alba (L.) Hassk. Direct shoot bud induction was observed on MS fortified with Kn (2 mg/l). However, maximum number of shoots was achieved when GA3, 0.5 mg/l was added to induction medium along with 1?M copper sulphate (ten times the normal MS level). Optimization of nutrient level in the basal medium promoted maximum regeneration response from both shoot tips and nodal explants. Elongated shoots were rooted in MS supplemented with IBA, 1.0 mg/l. Healthy, green plantlets with well developed roots, flowered normally in the field. Genetic stability of micropropagated plantlets was evaluated using RAPD markers. The amplification products were monomorphic in micropropagated plantlets and similar to those of mother plant revealing the genetic uniformity of plantlets. The regeneration protocol is highly efficient and reproducible so would be useful for mass multiplication, ex situ conservation and genetic transformation of E. alba (L.) Hassk.Plant Tissue Cult. & Biotech. 24(2): 223-234, 2014 (December)

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