Micropropagation and assessment of genetic stability in Celastrus paniculatus: An endangered medicinal plant

Biologia ◽  
2013 ◽  
Vol 68 (4) ◽  
Author(s):  
Sunil Senapati ◽  
Subhashree Aparajita ◽  
Gyana Rout

AbstractA highly efficient protocol for in vitro regeneration of an indigenous, endangered medicinal plant Celastrus paniculatus was achieved using nodal explants. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg/L 6-benzylaminopurine (BAP) and 0.1 mg/L naphthaleneacetic acid (NAA) showed maximum percentage of shoot multiplication (83.4%) with 8.2 shoots/explants. Maximum rooting of 73.3% with 4.8 roots/shoot was achieved on half-strength MS media supplemented with 0.5 mg/L indole-3-acetic acid (IAA) and the percentage of survival was 91% after acclimatization. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) marker study confirmed genetic stability for in vitro raised explants by showing 100% monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant species.

2008 ◽  
Vol 63 (9-10) ◽  
pp. 747-754 ◽  
Author(s):  
Sujata Mohanty ◽  
Manoj K. Panda ◽  
Enketeswara Subudhi ◽  
Laxmikanta Acharya ◽  
Sanghamitra Nayak

A protocol was developed for the in vitro propagation of ginger (Zingiber officinale) cv. Suprava using dormant axillary buds from unsprouted rhizomes. The dormant axillary buds embedded in the rhizome nodes were induced to sprout when cultured on MS medium supplemented with 6-benzyladenine (BA) alone (1 - 6 mg/l) or with a combination of BA (1 - 6 mg/l) and indole-3-acetic acid (IAA) (0.5, 1 mg/l). In vitro sprouted buds were transferred to the multiplication medium containing various combinations of auxins and cytokinins. MS basal medium supplemented with BA (1 mg/l), IAA (1 mg/l) and adenine sulfate (100 mg/l) was found optimum for the in vitro multiplication of shoots producing (8.2 ± 0.2) shoots from a single explant within 30 days of culture. The multiplication rate remained unchanged in subsequent subcultures. Rooting of shoots occurred in the same multiplication media. Upon transfer of the in vitro culture to ex vitro in pots, 96% of plants survived and established successfully under natural conditions. Tissue culture-raised plantlets of ginger could be conserved in vitro through subculturing at an interval of 4 months. The genetic stability of micropropagated clones was evaluated at regular intervals of 6 months up to 24 months in culture using cytophotometric estimation of 4C nuclear DNA content and random amplified polymorphic DNA (RAPD) analysis. Cytophotometric analysis revealed a unimodal distribution of the DNA content with a peak corresponding to the 4C value (23.1 pg), and RAPD analysis revealed monomorphic bands showing the absence of polymorphism in all fifty regenerants analyzed, thus confirming the genetic uniformity among in vitro grown somaclones of Z. officinale. This study is of commercial significance as axillary bud explants are available throughout the year for initiating a fresh culture of the elite ginger cv. Suprava to be used as a source of true-to-type disease-free planting material thereby minimizing the adverse effect of repeated subculturing from the same explant source.


1970 ◽  
Vol 18 (2) ◽  
pp. 173-179 ◽  
Author(s):  
T. Mallikadevi ◽  
P. Senthilkumar ◽  
S. Paulsamy

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)


2018 ◽  
Vol 135 (1) ◽  
pp. 111-118 ◽  
Author(s):  
Janhvi Mishra Rawat ◽  
Aakriti Bhandari ◽  
Susmita Mishra ◽  
Balwant Rawat ◽  
Ashok Kumar Dhakad ◽  
...  

1970 ◽  
Vol 16 (2) ◽  
pp. 85-94 ◽  
Author(s):  
S Karuppusamy ◽  
C Kiranmai ◽  
V Aruna ◽  
T Pullaiah

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)


2010 ◽  
Vol 19 (1) ◽  
pp. 71-78 ◽  
Author(s):  
A.K.M. Sayeed Hassan ◽  
Farhana Afroz ◽  
Miskat Ara Akhter Jahan ◽  
Rahima Khatun

A protocol was established for mass propagation of the valuable medicinal plant Ficus religiosa L. (Moraceae) through in vitro culture using apical and axillary buds of young sprouts from selected plants. Best shoot induction was observed on MS basal medium supplemented with 0.5 mg/l BAP + 0.1 mg/l IAA, in which 78 per cent of the explants produced 16 shoots per culture. Repeated subcultures in the same medium, resulted rapid shoot multiplication with 24 shoots per culture. In vitro raised shoots rooted on half strength MS supplemented with 2.0 mg/l IBA + 0.1 mg/l NAA. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for seven days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85 per cent.  Key words: Ficus religiosa, Medicinal plant, Shoot proliferation, Regeneration,                   Acclimatization D.O.I. 10.3329/ptcb.v19i1.4987 Plant Tissue Cult. & Biotech. 19(1): 71-78, 2009 (June)


Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 187
Author(s):  
Jorge Juan-Vicedo ◽  
Francisco Serrano-Martínez ◽  
Miriam Cano-Castillo ◽  
José Luis Casas

Tetraclinis articulata (Vahl) Masters is an endangered tree growing in coastal and arid environments that is widely exploited by the timber and resin industry, among other applications. In this context, the use of in vitro techniques is highly encouraged for its propagation. We present a protocol for micropropagation using twigs from adult trees as a source of explants. The Schenk and Hildebrandt basal medium (SH) supplemented with 30 g L−1 sucrose, 6.5 g L−1 plant agar, 4.0 mg L−1 6-benzyladenine (BA), and 0.05 mg L−1 1-naphthaleneacetic acid (NAA) provided the optimum multiplication rate (90.48 ± 9.52 explants with basal shoots and 2.58 ± 0.29 basal shoots per explant). Application of activated charcoal (AC) or ½ Knop solution in a liquid overlay produced significantly longer shoots. Supplementation of solid media with indole-3-butyric acid (IBA) or NAA gave low rooting percentages (<17%). Addition of 0.9 g L−1 AC improved rooting (40%) but rooting performance was optimal (66.7%) after a pulse treatment consisting of 4 h immersion in liquid SH medium without growth regulators, followed by 8 weeks of cultivation. Rooted microplants were successfully acclimatized (93.33%) in a peat moss and vermiculite mixture (1:1 v/v ratio). The genetic stability of the in vitro regenerated plantlets was confirmed using the randomly amplified polymorphic DNA (RAPD) technique. Explant survival and growth remained higher than 90% after 28 weeks of cold storage at both 4 °C and 10 °C. The protocol presented here allows for largescale T. articulata production and could be applied for both ex situ conservation strategies and industrial purposes.


2017 ◽  
Vol 9 (2) ◽  
pp. 50-70 ◽  
Author(s):  
Anusha Tharayil Koon Sasidharan ◽  
Megha Kizhakkepurakk Balachandr ◽  
Joseph Madassery ◽  
Kothanam Kuzhiyil Elyas

2004 ◽  
Vol 39 (2) ◽  
pp. 197-199 ◽  
Author(s):  
Rachel Fatima Gagliardi ◽  
Georgia Pereira Pacheco ◽  
Carlos Alberto Oliveira ◽  
Leonardo Alves Carneiro ◽  
José Francisco Montenegro Valls ◽  
...  

In vitro regeneration of Arachis retusa was examined for the purpose of germplasm renewal and conservation. Random amplified polymorphic DNA (RAPD) fingerprinting was used to evaluate the genetic stability of plants derived from embryo axes and apical segments. Ten arbitrary decamer primers were screened and five of them were selected. Ninety genomic regions were evaluated, with an average of 18 loci per clone. All amplified segments were monomorphic. The results indicate that recovered plants are genetically stable at the assessed genomic regions and that both regeneration processes are suitable for in vitro germplasm preservation of Arachis species.


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