Indirect regeneration of Ficus carica by the TCL technique and genetic fidelity evaluation of the regenerated plants using flow cytometry and ISSR

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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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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Double Haploid Development and Assessment of Androgenic Competence of Balkan Pepper Core Collection in Bulgaria

Plants ◽

10.3390/plants10112414 ◽

2021 ◽

Vol 10 (11) ◽

pp. 2414

Author(s):

Stanislava Grozeva ◽

Gancho Pasev ◽

Vesela Radeva-Ivanova ◽

Velichka Todorova ◽

Valentina Ivanova ◽

...

Keyword(s):

Flow Cytometry ◽

Double Haploid ◽

Genetic Improvement ◽

Core Collection ◽

Culture Medium ◽

Doubled Haploids ◽

Caps Marker ◽

Regenerated Plants ◽

Direct Embryogenesis ◽

Pepper Breeding

This study was designed to assess the androgenic potential of 180 pepper accessions and 11 progenies (four F1 and seven BC) possessing PMMoV resistance in order to complement an ongoing pepper breeding program. The experiment was carried out in 10 replications with 20 anthers for each accession in two different induction mediums from 2017 to 2019. The highest androgenic response was observed in culture medium 17-2 but differences between two mediums were nonsignificant. From a total of 191 genotypes, 102 genotypes expressed a potential for direct embryogenesis. Embryo induction was seen to be genotype-dependent and decreased in the following order: Pumpkin > Conical > Bell or blocky > Round > Elongate as the most responsive genotypes with over 10% reacted anthers being observed in CAPS-23, CAPS-29, CAPS-127, CAPS-157, CAPS-169, F1 and BC 887 derived from CAPS-23. The number of regenerated plants was higher in the conical group and least in the round varietal group. Regenerated plants were examined visually and by flow cytometry for identification of spontaneous doubled haploids (DH) and haploids. Those originating from F1 and BC progenies were additionally evaluated by a CAPS marker targeting L4 allele for resistance against PMMoV. Obtained results revealed two groups consisting of homozygous susceptible and resistant plants. Therefore, use of anther culture in ongoing breeding will greatly facilitate the pepper genetic improvement.

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

International Journal of Current Microbiology and Applied Sciences ◽

10.20546/ijcmas.2018.709.146 ◽

2018 ◽

Vol 7 (09) ◽

pp. 1223-1231 ◽

Cited By ~ 1

Author(s):

Asm ita ◽

S.S. Sindhu ◽

M. Jayanthi ◽

M.R. Dhiman ◽

M.K. Singh ◽

...

Keyword(s):

Genetic Fidelity ◽

Regenerated Plants

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Factors affecting in vitro regeneration of Ficus carica L. and genetic fidelity studies using molecular marker

Journal of Plant Biochemistry and Biotechnology ◽

10.1007/s13562-020-00590-9 ◽

2020 ◽

Author(s):

M. Moniruzzaman ◽

Zahira Yaakob ◽

Nurina Anuar

Keyword(s):

Molecular Marker ◽

In Vitro Regeneration ◽

Genetic Fidelity ◽

Ficus Carica ◽

Factors Affecting

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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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