In Vitro Propagation of Murraya koenigii L. Spreng (Curry Leaf Plant) through Adventitious Shoot Proliferation from Internode Explants (original) (raw)
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Micropropagation of Betula Pendula Roth Cultivars by Adventitious Shoot Induction from Leaf Callus
Acta horticulturae, 2010
Fifty-year-old specimens of Betula pendula 'Dalecarlica', 'Fastigiata', 'Purpurea', 'Youngii', and var. typica were used as donor plants. For callus induction, leaves obtained from in vitro were used as initial explants. The effects of the cultivar types and different media (MS, WPM and S) supplemented with zeatin (2, 5, and 10 mg L -1 ) or BA (0.5, 0.8, and 1.0 mg L -1 ) on adventitious bud formation from callus segments were studied. The best regeneration potential was observed in 'Youngii' and the purest in 'dalecarlica'. The best results were obtained on medium S containing 5 and 10 mg L -1 zeatin. The effect of IBA (0.3 and 0.5 mg L -1 ), NAA (0.3 and 0.5 mg L -1 ) and their combinations on the rooting of adventitious shoots was investigated. The highest percentage of rooting (100%) and the best root system was achieved on half-strength MS medium in combination with 0.3 mg L -1 IBA and 0.3 mg L -1 NAA. The high air humidity and the type of substrate were important factors for successful acclimatization. Maximum survival of plants (from 98.3 ± 1.7% up to 100.0 ± 0.0%) was obtained by opening the test tubes in the cultivation room for a period of 6 days, ensuring high air humidity for 14 days, and utilizing peat. The genotypes of the donor and in vitro propagated plants were analyzed by seven isozyme systems (GOT, IDH, LAP, MDH, PGI, PGM, and 6-PGD). Changes of the isozyme fractions of in vitro-propagated plants in comparison with the donor plants were not found.
In vitro Plant Propagation: A Review
2011
Micropropagation is an alternative mean of propagation that can be employed in mass multiplication of plants in relatively shorter time. Recent modern techniques of propagation have been developed which could facilitate large scale production of true-to-type plants and for the improvement of the species using genetic engineering techniques in the next century. An overview on the in vitro propagation via meristem culture, regeneration via organogenesis and somatic embryogenesis is presented. The usefulness of the plants in commercial industry as well as propagation techniques, screening for various useful characteristics and the influence of different cultural conditions in the multiplication, rooting and acclimatization phases on the growth of tissue cultured plant discussed.
International Journal of Science and Research (IJSR), 2015
An in vitro plant regeneration system was developed via direct somatic embryogenesis from different seedling explants of an important medicinal plant Murraya koenigii (L.) Spreng Cotyledons (COT), Hypocotyl (HYP) (10 to 15 mm) and Root (RT) segments (10 to 20 mm) were excised from 60 days old seedling as explants. The somatic embryos induction was achieved on Murashige and Skoog (MS) basal medium augmented with different concentrations of 6-benzyleaminopurine (BAP) 1.33 to 8.40 μM and thidiazuron (TDZ) 1.08 to 9.82 μM. The globular embryos originated from cut ends and entire surface of the root, hypocotyl explants and margins of cotyledons within 30-40 days. The percentage of somatic embryos induction per explant was significantly higher in HYP explants (94.21±5.77%) in the MS basal medium supplemented with 6.20 μM BAP and 8.64 μM TDZ. The highest rate of conversion of torpedo, heart and cotyledonary stages from globular stage was obtained in MS medium supplemented with 8.64 μM TDZ. The matured somatic embryos were transferred to the MS basal medium without Plant Growth Regulators (PGRs). Highest 88% of the matured embryos were germinated on transfer to ½ MS basal medium without PGR, where they grew for a further 3-4 weeks. Out of seventy six hardened plants seventy (92%) plantlets were found healthy under field conditions.
In vitro propagation of Murraya koenigii by axillary bud proliferation using mature explants
2018
Murraya koenigii is an important medicinal and aromatic plant. This plant is a native species of arid and semi-arid area of Rajasthan, India. Its leaves used as an analgesic, astringements, anti-dysenteric, antioxidant, hypo-lipidemic and for regulation of fertility. An efficient and reproducible procedure for the large scale propagation of M. koenigii is described. High frequency axillary shoot proliferation was induced in mature nodal explants. Regeneration of shoots (8-10) achieved on Murashige & Skoog (MS) medium (liquid) supplemented with benzylaminopurine (11.09 μM), kinetin (11.61 μM) and adenine sulphate (81.44 μM). Rooting (95%) in regenerated shoots occurred in 3 4 weeks of transfer on MS basal medium containing 12.30 μM indole butyric acid (IBA) and 5.13 μM indole acetic acid (IAA). In vitro plantlets regenerated from axillary bud proliferation were hardened for four weeks in a green house. The hardened plantlets were transferred to field conditions and high (82 to 85%) s...
Effects of Different Culture Media and Plant Growth Regulators on Micropropagation of ʻgisela 5ʼ
2020
We evaluated the influence of various culture media and plant growth regulators (PGR) upon axillary shoot proliferation rates and shoot lengths obtained in the in vitro multiplication stage in cherry rootstock 'Gisela 5'. The treatments consisted of the use of three basal media: Driver and Kuniyuki (DKW), Murashige and Skoog (MS), Woody Plant Medium (WPM) as well as three plant growth regulators (PGR) in various concentrations and combinations: N6-benzyladenine (BA at 0.3 and 0.5 mg/l), DL-Dihydrozeatin (DHZ at 1.0 and 2.0 mg/l) and N-benzyl-9-(tetrahydropyranyl)-adenine (BPA at 1.0 mg/l). The results show that the type of basal media and PGRs influenced the quantity as well as the quality of axillary shoot development. In the presence of BA at 0.3 and 0.5 mg/l in DKW media the proliferation rates (PR) were superior to those in MS and WPM media, both at 0.3 mg/l BA (PR = 6.66 ± 0.65) and 0.5 mg/l BA (PR = 8.93 ± 0.86) in the aforementioned DKW treatments. The DKW culture med...
In vitro propagation of Althaea officinalis : the role of plant growth regulators in morphogenesis
BioTechnologia
Althaea officinalis L. (marshmallow) belonging to the Malvaceae family, is an important plant that contains a variety of important phytocompounds including asparagine, pectin, flavonoids, polyphenolic acid, and scopoletin. The yield of these compounds can be improved using biotechnological methods that allow for a steady and continuous regeneration of plant material. To the best of our knowledge, thus far, the in vitro clonal multiplication of marshmallow has not been attempted on a large scale. Therefore, in this study, we developed callus induction and multiple shoot regeneration protocols from explants. All the explants, i.e., roots, nodes, and leaves, evoked compact white or yellow calli in a medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), which grew vigorously. The callus induction frequency was the highest (62.1%) from stem nodes, followed by leaves (39.1%) and roots (27.5%). The differential behavior of explants in response to various plant growth regulators (PGRs) was studied. The calli from leaves and roots were noted to be non-organogenic/embryogenic in media containing different PGR concentrations and have been described in this communication. The stem nodes used were cultured on MS media amended with different concentrations of benzyl-amino-purine (BAP: 0.5, 1.0, and 2.0 mg/l). Multiple shoots were formed at variable numbers, the maximum being in a medium supplemented with 1.0 mg/l of BAP. The induced shoots were rooted in IBA-, NAA-, and IAA-amended media, where IBA at 0.5 mg/l induced a maximum number of roots (8.8 roots/shoot). The regenerated plants were transferred to plastic pots, filled with soilrite and soil (1 : 1), and finally, transferred to outdoor conditions.
Bambusa nutans subsp. cupulata, a perennial tall grass bamboo has hig h potential value for economic and social aspects. Due to unpredictable flowering and seeds, it is obligated to propagate vegetative ways but it is inefficient and costly as well as the destruction of the huge stock of bamboo plants. The alternative method , micropropagation enhances a large-scale production of it. For this, the nodal segment was inoculated in MS solid and liquid media with a supplement of different concentrations of growth hormones. This article is reported the optimized in vitro propagation protocol to regenerate the large scale of the plants. The maximum bud sprout was observed in liquid (100%) with 2.5±0.19 cm length of shoots and solid (98.67%) with 3±0.19 cm length of shoots in MS media supplemented with of BAP (1mg/L) and KN (0.5 mg/L) in combined. The multiplication of shoots rapidly occurred on the liquid media fortified with BAP (1mg/L) and KN (0.5 mg/L). In vitro rooting was well achieved (84.67%) in the MS liquid media than solid media with fortified combined auxins 3 mg/L NAA and 3 mg/L IBA. We obtained 92.80% in primary (15 days) and secondary (3 months) hardening which yields a 100% survival rate in field transfer. We optimized the simple protocol for in vitro propagation of B. nutans subsp. cupulata. An efficient, but simple protocol, which can be applied for mass multiplication of B. nutans subsp. cupulta plant through nodal segment using the in vitro propagation technique.
In Vitro Cellular & Developmental Biology - Plant, 2008
A rapid and efficient method for the large-scale propagation of an endemic medicinal plant, Andrographis neesiana Wight, through in vitro culture of nodal explants obtained from 30-d-old aseptic seedling has been developed. High frequency direct shoot proliferation was induced in nodal explants cultured on Murashige and Skoog's medium supplemented with thidiazuron. Amongst the various cytokinins tested (BAP, kinetin, thidiazuron and 2-isopentyl adenine), thidiazuron proved to be the most effective. The shoot forming capacity of the nodal explants was influenced by the thidiazuron concentration (1-12.5 µM) and the optimal response was observed at 10 µM thidiazuron, which induced an average of 34 shoots in 94% of the cultures within 4 wk. Significant differences were recorded in terms of average number of shoots per explant (8.6-34.1) among the different concentrations of thidiazuron investigated. Concentrations of all cytokinins tested reach a level that can be considered above the optimum level, as marked by a reduced frequency of shoot proliferation. The multiple shoots obtained on various concentrations of thidiazuron failed to elongate even after transfer to hormone-free MS medium. Elongation of the induced shoots was achieved on MS basal medium supplemented with 1.0 µM GA within 2 wk. A proliferating shoot culture was established by repeatedly 3 subculturing the original nodal explants on shoot multiplication medium after each harvest of the newly formed shoots. The explants retained their morphogenic potential even after three harvests. Therefore, in 90 d, about 60-70 shoots were obtained from a single nodal explant and the nodal explants from primary shoots further regenerated equivalent number of shoots, depicting their high frequency regeneration potential in A. neesiana. Rooting was best induced in 94% of shoots cultured on MS medium supplemented with 2.5 µM indole-acetic acid (IAA), within a wk. The plantlets were successfully transferred to soil after hardening with a 92% survival rate. The system is rapid: the initiation of shoot buds to the transplanting of regenerants to soil is completed in 8-9 wk.
A micropropagation protocol for Cassia angustifolia Vahl. from root explants
Acta Physiologiae Plantarum
The aim of this study was to develop a new micropropagation system for Cassia angustifolia Vahl., an important medicinal legume using root explant as starting material. Root explants taken from 30-day-old aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators: 6-benzyladenine (BA), kinetin (Kn), and thidiazuron (TDZ). Organogenic nodular calli obtained on MS ? TDZ (1.0 lM) were transferred to shoot regeneration medium supplemented with different cytokinins (BA, Kn or TDZ) either alone or in combination with auxin:indole-3-acetic acid or a-naphthalene acetic acid. Maximum shoot regeneration frequency (90%) was obtained on MS ? BA (2.5 lM) ? NAA (0.6 lM) wherein a maximum of 42.76 ± 1.47 shoot buds per explant were induced with a maximum conversion rate of 35.63 ± 0.75 shoots per explant and average shoot length of 5.43 ± 0.20 cm. Elongated microshoots were successfully rooted under ex vitro conditions by pulse treatment in 200 lM of indole-3butyric acid for half an hour. Microshoots were rooted, acclimatized and hardened off simultaneously in sterilized soilrite inside the growth room and then established in pots containing sterilized soil and manure (1:1) and grown under greenhouse condition with 90% survival rate. The histological sections at different developmental stages of shoot buds revealed the organization of nodular meristematic zone leading to the orientation and differentiation of shoot buds in large number and thereafter conversion into healthy shoots.
In vitro regeneration through adventitious buds in
High frequency shoot regeneration from in vitro derived leaf explants of Wattakaka volubilis (L.f.) Stapf was achieved through callus mediated organogenesis. Organogenic calli were induced from 20 day old aseptic seedling explants on Murashige and Skoog medium fortified with various concentrations and combinations of plant growth regulators, benzylaminopurine (BAP), α-naphthaleneacetic acid (NAA), indole 3-butyric acid (IBA) and gibberellic acid (GA3). A mean of 8.6 shoots developed from organogenic callus induced from a 2 x 2 cm leaf explants on MS medium with 3% sucrose having 5.37μM NAA in combination with 2.22 μM BAP with 60% induction capacity. Further development of adventitious shoots could be achieved by sub culturing the callus to the same medium with 4.40 μM BAP and 0.288 μM GA3. Organogenesis could not be achieved from the calli of ex vitro derived leaf explants. The developed shoots were rooted on half-strength MS medium with 1% sucrose, 4.90 μM IBA and 0.93 μM kinetin at a frequency of 85%. Well rooted plantlets were then transplanted to vermiculite soil (3:1) mixture in polythene covered pots kept under culture room conditions. Approximately 60% plantlets survived and grew into whole plants.