scholarly journals Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Gulab Khan Rohela ◽  
Phanikanth Jogam ◽  
Prasad Bylla ◽  
Christopher Reuben
Keyword(s):  
Medicinal Plant ◽  
Rapd Marker ◽  
Genetic Fidelity ◽  
Mother Plant ◽  
Garden Soil ◽  
Naphthalene Acetic Acid ◽  
Stem Explants ◽  
Clonal Multiplication ◽  
Indirect Regeneration

Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.

scholarly journals MICROPROPAGATION OF AN ENDANGERED AND ENDEMIC MEDICINAL PLANT CAYRATIA PEDATA VAR. GLABRA

2020 ◽  
pp. 191-196
Author(s):  
SHARMILA S ◽  
RAMYA EK ◽  
MOWNIKA S ◽  
DHIVYA SM
Keyword(s):  
Medicinal Plant ◽  
Callus Formation ◽  
Naphthalene Acetic Acid ◽  
Stem Explants ◽  
Root Induction ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Mass Multiplication ◽  
Endemic Medicinal Plant

Objective: The objective of the present study was to develop standardization protocol for the successful in vitro mass propagation of Cayratia pedata var. glabra through leaf and stem explants, since it is a rare, endangered, and endemic medicinal plant using biotechnological involvements and to conserve this endangered species. Methods: The application of biotechnological principles for the establishment of micropropagation under in vitro conditions has been studied by following the methods. The explants, namely, leaf and stem harvested from in vivo plants were thoroughly washed and properly sterilized with sterilients. The explants were transferred to Murashige and Skoog (MS) medium supplemented with growth regulators 6-benzyladenine (BAP) and naphthalene acetic acid (NAA) in the concentration range of 0.5–3.0 mg/l which were tested for callus induction and morphogenesis. The elongated shoots were transferred to MS medium supplemented with NAA at different concentrations for root induction. Results: The explants collected from the field (shola) were treated in different steriliants with various concentrations at different time for sterilization. Among the various combinations tried, the Teepol treatment for 10 min followed by bavistin 20 min, antibiotics, namely, ampicillin and rifampicin for 20 min, 70% alcohol for 30 s, and 0.12 % HgCl2 for 3 min was found to be effective. The explants were cultured in MS medium supplemented with various concentrations of BAP and NAA. The results noted that an increase in the concentration of BAP concomitantly reduced the frequency of callus formation. The maximum callusing frequency and more number of shoot formation was observed in the lower concentration of BAP (0.5 mg/l) in combination with NAA (0.2 mg/l). The callus obtained from all the above combinations was sub-cultured on MS medium with same combinations of BAP and NAA. The maximum frequency of root formation in leaf callus was 85% and 75% in stem callus and both were achieved on MS medium with NAA (1 mg/l) after 2 weeks. Conclusions: The current investigation provides a competent in vitro propagation method for C. pedata var. glabra which could be commercialized for developing identical plants with high-quality mass multiplication rate and for better conservation of the germplasm. Both the methods described here are well suited for the mass multiplication of this critically endangered and endemic climber species.

scholarly journals In Vitro Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey

2020 ◽  
Author(s):  
Nurşen Çördük ◽  
Cüneyt Aki
Keyword(s):  
Acetic Acid ◽  
Medicinal Plant ◽  
In Vitro Propagation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Northwestern Turkey ◽  
Helen Of Troy ◽  
Endemic Medicinal Plant

Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regeneration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber.

scholarly journals In vitro Regeneration of the Medicinal Plant, Plumbago zeylanica L. with Reference to a Unique Population in Maruthamalai, the Western Ghats, India

1970 ◽  
Vol 18 (2) ◽  
pp. 173-179 ◽  
Author(s):  
T. Mallikadevi ◽  
P. Senthilkumar ◽  
S. Paulsamy
Keyword(s):  
Medicinal Plant ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Garden Soil ◽  
Plumbago Zeylanica ◽  
Adventitious Shoot Formation ◽  
The Western Ghats

The in vitro regeneration of Plubago zeylanica exhibited that the callus was initiated in the basal medium containing BAP, NAA, 2, 4-D, and IBA.  The high amount (90%) of organic calli was induced in the basal medium supplemented with 2, 4-D, alone at 2.0 mg/l. In the subculture the adventitious shoot formation was prominently higher (83%) in the basal medium containing BAP, and NAA at 3.5 and 0.3 mg/l, respectively. IAA (1.0 mg/l)effectively produced higher percen-tage (90) of roots and root growth. After sequential hardening, survivability rate was observed to be significantly higher (80%) in the hardening medium containing garden soil, sand and vermicompost in the ratio of 1 : 1 : 1 by volume under greenhouse condition.  Key words: Plumbago zeylanica, In vitro regeneration, Medicinal plant D.O.I. 10.3329/ptcb.v18i2.3648 Plant Tissue Cult. & Biotech. 18(2): 173-179, 2008 (December)

scholarly journals Micropropagation of Vanasushava pedata - An Endangered Medicinal Plant of South India

1970 ◽  
Vol 16 (2) ◽  
pp. 85-94 ◽  
Author(s):  
S Karuppusamy ◽  
C Kiranmai ◽  
V Aruna ◽  
T Pullaiah
Keyword(s):  
Medicinal Plant ◽  
In Vitro Regeneration ◽  
Continuous Production ◽  
Natural Habitat ◽  
Nodal Segments ◽  
Garden Soil ◽  
Axillary Shoot ◽  
Similar Frequency ◽  
Endangered Medicinal Plant

An efficient in vitro propagation of an endangered medicinal plant Vanasushava pedata (Apiaceae) by axillary shoot proliferation from nodal segments of mature plants was designed. The medium type and growth regulators markedly influenced in vitro regeneration of V. pedata. An in vitro plantlet production system has been investigated on MS with the synergistic combination of BA (5.0 mg/l), IAA (0.1 mg/l) and 3 % sucrose which promoted the maximum number of shoots (8.6) as well as enhanced shoot lengths. Subculturing of nodal segments from in vitro derived shoots on a similar medium enabled continuous production of healthy shoots with a similar frequency. Rooting was highest (100%) on half strength MS containing IAA (2.0 mg/l). Micropropagated plants established in garden soil and forest humus (1 : 1) were uniform and identical to the donor plants with respect of growth characteristics as well as floral features. These in vitro-raised plants grew normally in greenhouse and natural habitat without showing any morphological variation.  Key words: Vanasushava pedata, Medicinal plant, Nodal explants, Micropropagation, Successful acclimationDOI = 10.3329/ptcb.v16i2.1109Plant Tissue Cult. & Biotech. 16(2): 85-94, 2006 (December)

scholarly journals Indirect in vitro Regeneration of the Medicinal Plant, Aspilia africana, and Histological Assessment at Different Developmental Stages

2021 ◽  
Vol 12 ◽  
Author(s):  
Denis Okello ◽  
Sungyu Yang ◽  
Richard Komakech ◽  
Yuseong Chung ◽  
Endang Rahmat ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Developmental Stages ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Root Explants ◽  
Indirect Regeneration

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.

scholarly journals Micropropagation and Genetic Fidelity of the Regenerants of Aglaonema ‘Valentine’ Using Randomly Amplified Polymorphic DNA

HortScience ◽  
2016 ◽  
Vol 51 (4) ◽  
pp. 398-402 ◽  
Author(s):  
Mohammed Elsayed El-Mahrouk ◽  
Yaser Hassan Dewir ◽  
Yougasphree Naidoo
Keyword(s):  
In Vitro Regeneration ◽  
Shoot Proliferation ◽  
Genetic Fidelity ◽  
Naphthalene Acetic Acid ◽  
Axillary Shoot ◽  
Ms Medium ◽  
Randomly Amplified Polymorphic Dna ◽  
Simple Protocol ◽  
Regenerated Plantlets

The present study reports a simple protocol for in vitro regeneration of Aglaonema ‘Valentine’ using axillary shoot explants for rapid multiplication and production of true-to-type plants. Different concentrations of benzyladenine (BA; 0, 1, 3, 5, and 7 mg·L−1), kinetin (Kin; 0, 1, 3, 5, and 7 mg·L−1), thidiazuron (TDZ; 0, 0.5, 1.0, 1.5, and 2.0 mg·L−1), naphthalene acetic acid (NAA; 0, 0.5, and 1.0 mg·L−1), and indole-3-butyric acid (IBA; 0, 0.5, and 1.0 mg·L−1) were used for shoot regeneration. The highest shoot proliferation (5.0) was obtained on Murashige and Skoog (MS) medium supplemented with 1.5 mg·L−1 TDZ and 1 mg·L−1 NAA. In vitro rooting was easily achieved with 100% at all concentrations of NAA and IBA supplemented to half- or full-strength MS medium. Regenerated plantlets were acclimatized in greenhouse with 100% survival rate. Randomly amplified polymorphic DNA (RAPD) analysis confirmed the genetic fidelity of the regenerated plantlets and mother plant.

scholarly journals Impact of Hormones on the Proliferation of Shoots and Initiation of Roots in Salvia santolinifolia (Boiss), A High Value Medicinal Herb

2020 ◽  
Vol 63 (1) ◽  
pp. 30-36
Author(s):  
Tour Jan ◽  
Beena Naqvi ◽  
Ali Hazrat ◽  
Raiha Qadri ◽  
Muhammad Nisar ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Medicinal Plant ◽  
Butyric Acid ◽  
Medicinal Herb ◽  
In Vitro Conservation ◽  
Nodal Explants ◽  
Naphthalene Acetic Acid ◽  
Culture Age ◽  
Number Of Shoots

Salvia santolinifolia is a medicinal plant and an efficient in vitro conservation system is established. The influence of N6Benzylaminopurine (BAP), N6-(2-isopentyl)-adenine (2iP) and Kinetin at various concentrations were evaluated, single and in mixture with NAA (Naphthalene acetic acid) for the production of auxiliary shoots from nodal explants of S. santolinifolia. BAP at 3.0 mg/L in MS1 media produced maximum (11.66±3.38) number of shoots while elongated (5.37±1.45) shoots were produced in the MS2 medium in subcultures at 2.0 mg/L of 2iP. Least number of shoots were formed when auxin and cytokinin were used in combination. Length of culture, age was an important consideration for the initiation and development of roots. Rooting of shoot was attained with Indol-3-butyric acid (IBA) (3.0 mg/L) from shoots of 4th, 5th and 6th subculture while shoots taken from 1st, 2nd and 3rd subculture failed to form roots.  

scholarly journals MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

2019 ◽  
Vol 6 (1) ◽  
pp. 1-7
Author(s):  
Diego Pandeló José ◽  
José Marcello Salabert De Campos ◽  
Lyderson Facio Viccini ◽  
Emilly Ruas Alkimim ◽  
Marcelo De Oliveira Santos
Keyword(s):  
Flow Cytometry ◽  
Medicinal Plant ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Naphthalene Acetic Acid ◽  
Flow Cytometry Analysis ◽  
Ploidy Levels

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.

Sign in / Sign up

Export Citation Format

Share Document