scholarly journals Efficient In Vitro Propagation by Ex Vitro Rooting Methods of Artemisia absinthium L., an Ethnobotanically Important Plant

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Anthropogenic Activities ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Ex Vitro ◽  
Artemisia Absinthium ◽  
Genetic Stock ◽  
In The Wild ◽  
Important Medicinal Plant

Artemisia absinthium is an important medicinal plant. Owing to the increasing anthropogenic activities and demand from the pharmaceutical industry, this plant species is overexploited; thereby this endangered its genetic stock in the wild. Therefore, it is urgently needed to develop nonconventional methods for conservation of A. absinthium. Nodal segments obtained from the field grown 2-month-old plants were used as explants. Murashige and Skoog (MS) medium containing 0.5 mg/L 6-benzylaminopurine (BAP) and 0.25 mg/L kinetin (Kn) were reported to be optimum for induction of shoots (6.0 ± 0.52 shoots per explant). The shoots were multiplied by repeated transfer of original explants and by subculturing of in vitro raised shoots on MS medium augmented with 1.0 mg/L each of BAP and Kn and 0.1 mg/L α-naphthaleneacetic acid (NAA). All in vitro regenerated shoots (100%) were rooted (4.4 ± 0.35 roots) on one-fourth strength MS medium supplemented with 2.0 mg/L indole-3 butyric acid (IBA). Cent percentage shoots rooted ex vitro on sterile Soilrite under the greenhouse conditions when the shoots were treated with 200 mg/L of IBA for 5 min. Plantlets rooted in vitro and ex vitro were acclimatized successfully in the greenhouse and exhibited 87% and 95% survival rate.

scholarly journals In Vitro Regeneration, Ex Vitro Rooting and Foliar Stoma Studies of Pseudostellaria heterophylla (Miq.) Pax

Agronomy ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 949
Author(s):  
Fengyun Wang ◽  
Xiaowei Xin ◽  
Hao Wei ◽  
Xiaohui Qiu ◽  
Boling Liu
Keyword(s):  
Stomatal Density ◽  
In Vitro Regeneration ◽  
Wild Plants ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Ex Vitro ◽  
Pseudostellaria Heterophylla ◽  
Stomatal Apparatus

Pseudostellaria heterophylla, in the family Caryophyllaceae, is an important Chinese medicinal plant commonly used to treat various diseases in children and valued for its ornamental properties. In this study, nodal segments were obtained from wild plants and used as explants to develop an efficient micropropagation protocol for this species. Murashige and Skoog (MS) medium supplemented with 1.5 mg·L−1 6-benzyladenine (6-BA) was the most suitable medium for inducing axillary buds and enhancing their growth, and MS medium containing 0.1 mg·L−1 indole-3-butyric acid (IBA) was the most effective for inducing in vitro rooting. To reduce labor, time, and cost, microshoots were rooted under ex vitro conditions. Pretreatments of the shoots with 100 mg·L−1 naphthaleneacetic acid (NAA) for 1 min ensured successful rooting in 86.7% of shoots. Comparison of the leaf microstructure between in vitro- and ex vitro-rooted plantlets revealed abnormal stomatal apparatus in the former. The stomatal apparatus of ex vitro plantlets were normal, although the stomatal density was reduced, which indicated that these plantlets were more likely to be able to adapt to environmental conditions in the field. We identified the optimal medium for P. heterophylla multiplication with respect to increased rooting efficiency of micropropagated shoots under ex vitro conditions. This results presented here will be helpful for agricultural cultivation of P. heterophylla.

scholarly journals In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals Regeneration of Plants from Apical Meristem Tips and Nodal Segments of Arachis pintoi

2003 ◽  
Vol 30 (2) ◽  
pp. 75-79 ◽  
Author(s):  
H. Y. Rey ◽  
L. A. Mroginski
Keyword(s):  
Apical Meristem ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Potential ◽  
Arachis Pintoi ◽  
Solid Media ◽  
One Step

Abstract The in vitro regeneration potential of shoot apical tips (2 to 3 mm in length), meristems (0.3 to 0.5 mm in length), and nodal segments (4 to 7 mm long with an axillary bud) of diploid (2n = 2x = 20) and triploid (2n = 3x = 30) cytotypes of Arachis pintoi was evaluated. Explants were cultured on MS medium supplemented with different concentrations and combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA). In one experiment the effect of gibberellic acid was tested. The cultures were done in liquid and solid media. Plant regeneration can be readily achieved from all explants in one step of 30 d culture on MS + 0.01 mg/L each of NAA and BA or two steps consisting of 1) shoots regeneration through culture of explants on MS + 0.01 mg/L each of NAA and BA, and 2) induction of rooting in regenerated shoots by reculture on MS + 0.01 mg/L NAA. The plantlets were successfully transferred to pots in a greenhouse.

scholarly journals In vitro study of Tinospora cordifolia (Willd.) Miers (Menispermaceae)

2010 ◽  
Vol 6 ◽  
pp. 103-105 ◽  
Author(s):  
Aditi Singh ◽  
Saroj K Sah ◽  
Aunji Pradhan ◽  
Sabari Rajbahak ◽  
Niran Maharajan
Keyword(s):  
Medicinal Plant ◽  
Callus Induction ◽  
Shoot Growth ◽  
Tinospora Cordifolia ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Nodal Segment ◽  
Root Induction ◽  
Ms Medium

In vitro study was carried out in an important medicinal plant Tinospora cordifolia (Willd.) Miers belonging to the family: Menispermaceae. Vegetative parts such as stem, leaf and nodal explants were excised from an elite in vivo grown mature plant and thereafter cultured on Murashige-Skoog (MS) medium supplemented with different hormonal concentrations for callus induction and organogenesis. Callus formation occurred from nodal segments, leaf and inter-node explants when planted on different combinations of hormones. Tinospora cordifolia showed response for in vitro shoot growth from the nodal segment. The best shoot growth was observed on MS medium supplemented with kinetin (1.5 mg/l). Similarly, the best result for root induction was obtained on MS medium supplemented with 6-benzylaminopurine (1.0 mg/l) and naphthaleneacetic acid (2.5 mg/l). Key-words: callus induction; explants; medicinal plant; MS medium; tissue culture.DOI: 10.3126/botor.v6i0.2918 Botanica Orientalis - Journal of Plant Science (2009) 6: 103-105

scholarly journals In Vitro Multiplication and NMR Fingerprinting of Rare Veronica caucasica M. Bieb

Molecules ◽  
2021 ◽  
Vol 26 (19) ◽  
pp. 5888
Author(s):  
Desislava I. Mantovska ◽  
Miroslava K. Zhiponova ◽  
Milen I. Georgiev ◽  
Tsvetinka Grozdanova ◽  
Dessislava Gerginova ◽  
...  
Keyword(s):  
Ex Situ ◽  
Nodal Segments ◽  
Cultivated Plants ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Ex Vitro ◽  
Biotechnological Approach ◽  
Iridoid Glucosides ◽  
Development In Vitro

Micropropagation of rare Veronica caucasica M. Bieb. was achieved by successful in vitro cultivation of mono-nodal segments on MS medium supplemented with 1.0 mg L–1 6-benzylaminopurine (BA) and then transferring the regenerated plants on hormone free basal MS medium for root development. In vitro multiplicated plants were successively acclimated in a growth chamber and a greenhouse with 92% survival. The number of plastid pigments and the total phenolics content in in vitro cultivated and ex vitro adapted plants were unchanged, and no accumulation of reactive oxygen species (ROS) was detected by staining with 3-3′-diaminobenzidine (DAB) and 2′,7′-dichlorofluorescein diacetate (DCF-DA). Nuclear Magnetic Resonance (NMR) fingerprinting allowed for the identification of the major alterations in metabolome of V. caucasica plants during the process of ex situ conservation. Iridoid glucosides such as verproside, aucubin and catalpol were characteristic for in vitro cultivated plants, while in ex vitro acclimated plants phenolic acid–protocatechuic acid and caffeic acid appeared dominant. The successful initiation of in vitro and ex vitro cultures is an alternative biotechnological approach for the preservation of V. caucasica and would allow for further studies of the biosynthetic potential of the species and the selection of lines with a high content of pharmaceutically valuable molecules and nutraceuticals.

scholarly journals Plant Regeneration from Leaf Explants of the Medicinal Herb Wedelia chinensis

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Liver Toxicity ◽  
Leaf Explants ◽  
Basal Medium ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Average Percentage

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.

Micropropagation of two threatened Tasmanian species of Calocephalus (Asteraceae), with comments on phenotypic plasticity

2003 ◽  
Vol 51 (4) ◽  
pp. 415 ◽  
Author(s):  
Rebecca Jane Sands ◽  
Natalie Ruth Brown ◽  
Anthony Koutoulis
Keyword(s):  
Phenotypic Plasticity ◽  
Plant Growth Regulator ◽  
Root Formation ◽  
Indoleacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Ex Vitro ◽  
Morphological Differences

Micropropagation systems were developed for Calocephalus citreus Less. and C. lacteus Less., two threatened Tasmanian members of the Asteraceae. Disinfected cold-treated capitula were used to initiate regeneration. For C. citreus, initiation was achieved on Murashige and Skoog (MS) medium with 0.1�mg�L–1 or 0.5�mg�L–1 indoleacetic acid (IAA) and 1�mg�L–1 6-benzylaminopurine (BAP) in 5�weeks, while for C. lacteus initiation was achieved on MS with α-naphthaleneacetic acid (NAA) (0.1�mg�L–1) in 3�weeks and on MS without any plant growth regulator (PGR) in 6�weeks. Multiplication was achieved in both species on MS with various concentrations of IAA (0.01–0.5�mg�L–1) and BAP (0.1–1�mg�L–1). In C. citreus, shooting in all treatments did not differ significantly from PGR-free MS, while in C. lacteus PGR-free MS was one of the better treatments. Multiplication media also initiated root formation in C. lacteus, thereby facilitating immediate planting out. Optimal root induction in C. citreus was achieved by using MS with 1�g�L–1 activated charcoal. Clear morphological differences between in vitro and ex vitro plants of both species were observed. This phenotypic plasticity was more pronounced in C. lacteus than in C. citreus. As C. lacteus has a wider distribution than C. citreus and C. lacteus was more responsive during many stages of the micropropagation process, it may be possible to use the culture-induced phenotype to provide insights into the ecology of plant species.

scholarly journals Immature Embryo Germination and Its Micropropagation of Ilex crenata Thunb.

HortScience ◽  
2015 ◽  
Vol 50 (5) ◽  
pp. 733-737 ◽  
Author(s):  
Yujie Yang ◽  
Donglin Zhang ◽  
Zhihui Li ◽  
Xiaoling Jin ◽  
Jinying Dong
Keyword(s):  
Seed Germination ◽  
Germination Rate ◽  
Immature Embryo ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Embryo Germination ◽  
Average Survival ◽  
High Germination

To shorten Ilex seed germination time and speed up breeding cycles, immature embryos of Ilex crenata ‘Sky Pencil’ seedlings were removed from fruits at their heart-shape stage and cultured in vitro on Murashige and Skoog (MS) medium or Woody Plant Medium (WPM) with 3% sucrose and 0.65% agar. Cultures were incubated at 27 °C for 2 weeks in darkness and subsequently moved to a growth chamber with 14-hour photoperiod (115 μmol⋅m−2⋅s–1). Embryos began to germinate 2–3 weeks after culture. The highest germination rate was 91.67% under 1/4 MS medium. Embryos cultured on MS medium also had high germination rates and produced the longest seedlings to 8.02 mm. Nodal segments with one axillary bud taken from embryo germination seedlings were cultured on MS medium with various concentrations of cytokinins and auxins for micropropagation. Zeatin (ZT; 4-hydroxy-3-methyl-trans-2-butenylaminopurine) increased the number of shoots and shoot lengths significantly more than 6-benzylaminopurine (6-BA). The recommended ZT concentration should be 2.28 µM. Rooting induction could be established on 1/4 MS medium with various concentrations of indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA). IBA at 4.14 µM produced the best rooting percentage (91.67%) and good-root quality. All rooted plantlets were transplanted into a mixture of peatmoss and perlite (1:1 v/v) and acclimatized in a mist system. The average survival rate was 88.8%. The rapid embryo germination protocol for Ilex crenata could save Ilex breeders at least 2 years compared with traditional seed germination.

scholarly journals Micropropagation of Chinquapin (Castanea henryi) Using Axillary Shoots and Cotyledonary Nodes

HortScience ◽  
2018 ◽  
Vol 53 (10) ◽  
pp. 1482-1486 ◽  
Author(s):  
Huan Xiong ◽  
He Sun ◽  
Feng Zou ◽  
Xiaoming Fan ◽  
Genhua Niu ◽  
...  
Keyword(s):  
Adventitious Shoots ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Segments ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Cotyledonary Nodes

Castanea henryi is an important woody grain tree species native to China. The objective of the current study was to find the suitable plant growth regulators (PGRs) and the optimal concentrations for direct organogenesis by using axillary shoots and cotyledonary nodes. Seeds were collected from the field, sterilized, and germinated in vitro. Axillary shoots and cotyledonary nodes of 3-week-old seedlings were used as explants. To find the suitable PGR for adventitious shoot induction, 0.5 mg·L–1 6-benzylaminopurine (6-BA), 0.1 mg·L–1 indole-3-acetic acid (IAA), 0.1 mg·L–1 2,4-dichlorophenoxyacetic acid (2,4-D), or 0.1 mg·L–1 1-naphthaleneacetic acid (NAA) was supplemented to Murashige and Skoog (MS) medium containing 0.65% agar and 3% sucrose. A high induction percentage of adventitious shoots (85.67%) was obtained from cotyledonary nodes supplemented with 0.1 mg·L–1 2,4-D. The type of explant influenced shoot proliferation rates and quality. Apical explants produced more and longer shoots than nodal segments. For shoot multiplication, 1 mg·L–1 6-BA + 0.05 mg·L–1 indole-3-butyric acid (IBA) supplemented with MS medium produced 12.33 and 6.25 shoots per explant, respectively, from apical and nodal explants. For shoot elongation and strengthening, 2 mg·L–1 6-BA + 0.05 mg·L–1 IBA supplemented with MS medium was the best combination, producing shoots with a mean length of 3.50 cm, a diameter of 0.46 cm, and about eight leaves per shoot. The greatest rooting of 76.70% and 11.33 roots per shoot was achieved when cultured in MS medium supplemented with 3.5% perlite + 1.5 mg·L–1 IBA. For acclimatization of the rooted plantlets in the greenhouse, a survival rate of 80% was achieved. This protocol—from multiplication to acclimation—is helpful to realize mass propagation of high-quality trees of chinquapin for increasing production and nut quality.

scholarly journals Effects of Phytohormone and Regulators on Shoot Tip and Nodal Explants for In Vitro Shoot and Root Clonal Propagation of Vitex negundo L.

2021 ◽  
pp. 65-78
Keyword(s):  
Germplasm Conservation ◽  
Multiple Shoots ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Vitex Negundo ◽  
Surface Sterilization

Medicinal plants are one of the most vital natural resources, but many of them are currently endangered due to habitat loss. Consequently, it is critical to emphasize the importance of using micropropagation techniques for mass propagation of plantlets on a commercial scale, in addition to germplasm conservation and distribution. Nodal explants and shoot tips were expunged from 15 days of the explant by aseptic seedlings, an effective, quick, and better in vitro plant regeneration procedure for Vitex negundo L. has been developed. The recent study was considered to develop an in vitro procedure for the regeneration of V. negundo L., a traditional medicinal plant. Nodal segments and shoot tips were cultivated on MS medium enhanced with numerous plant growth regulators. For multiple shoots and root regeneration, various cytokinins were examined. 6-benzyl-aminopurin (BAP), kinetin (Kin), and 1H-indole-3-butanoic acid (IBA) were all tested as a supplement to Murashige and Skoog (MS) medium including auxin phytohormone, such as Indole acetic acid (IAA) and 1-naphthaleneacetic acid (NAA). The furthermost effective surface sterilization treatment for explants of V. negundo has been found 0.1% HgCl2 for 8 minutes. In all treatments, multiple shoots were collected from shoot tips and nodal segments. In MS media added with 2.0mg/l BAP, the most shoots were seen in V. negundo. Furthermore, V. negundo regeneration shoots rooted effectively in half MS containing 1.0 mg/l IBA. Finally, proliferated plantlets were effectively adapted in soil, where they grew normally without morphological anomalies and had a survival rate of 92 percent.

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