scholarly journals In-vitro Regeneration from Direct and Indirect Organogenesis of Crescentia alata Kunth an Important Multipurpose Medicinal Tree

Author(s):  
R. Abinaya

In this present work, an in-vitro regeneration protocol for Crescentia alata (C. alata) was developed using various explants on Murashige and Skoog (MS) medium augmented with different concentrations and combinations of plant growth regulators (PGRs) for direct and indirect regeneration. The direct organogenesis was established from nodes and internodes on MS medium supplemented with cytokinins and auxins. The indirect organogenesis via callus phase was obtained from leaf, nodes and internodes on MS medium supplemented with different concentrations of PGRs. The high frequency shoot organogenesis were achieved directly from nodal explants were cultured on MS medium supplemented with 3.0 mg/L BAP+0.5 mg/L KIN +1.0 mg/L NAA. Indirect organogenesis callogenic frequency was optimized at the concentration of MS medium containing 1.0 mg/L BAP + 5.0 mg/L IAA. The callus was obtained from all the explants were used, among these explants internodal explants gave best result on MS medium supplemented with different concentrations of cytokinins and auxins for indirect organogenesis experiment. Indirect organogenesis the highest number of shoot regeneration was obtained in MS Basal Medium with 4.0 mg/L BAP + 0.5 mg/L KIN + 2.0 mg/L NAA from internodal explants. For root formation the regenerative shoots which were sub cultured on MS medium containing different ratios of auxins. The rooted plantlets were transferred successfully to the pots containing sterilized soil and were successfully hardened at greenhouse condition for 20 days then exposed to the natural environment. This is the first successful micropropagation report of an efficient and rapid in-vitro clonal propagation protocol for C. alata by direct and indirect shoot organogenesis through various explants, which can be employed for conservation of this important medicinal tree species as well as the utilization of an biologically important active biomolecules. This protocol can be very useful to obtain plants from various explants, without the requirement of meristematic regions, enabling the obtainment of a higher number of plants in short period.

2008 ◽  
Vol 34 (No. 2) ◽  
pp. 84-89 ◽  
Author(s):  
R. Rout G ◽  
A. Mahato ◽  
K. Senapati S

Rapid shoot multiplication of Nyctanthes arbortristis was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0−1.5 mg/l 6-benzyladenine (BA), 50 mg/l adenine sulfate (Ads) and 3% (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA + Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on the MS medium supplemented with 1.5 mg/l BA, 50 mg/l Ads and 0.1 mg/l IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 days on ½ strength MS medium supplemented either with indole-3-butyric acid (IBA), IAA or naphtylacetic acid (NAA) with 2% sucrose. Maximum percentage of rooting was obtained on medium having 0.25 mg/l IBA, 0.1 mg/l IAA and 2% sucrose. About 70% of rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the soil condition. This result will facilitate the conservation and propagation of the important medicinal plant.


2014 ◽  
Vol 20 ◽  
pp. 99-108 ◽  
Author(s):  
MS Islam ◽  
MA Bari

Context: The application of encapsulated shoot tips and nodal segments may contribute to the protection of rare and threatened medicinal plants. Although the artificial seed technique has been reported for more than two decades, for medicinal plants this method has not been developed sufficiently. The main limitations in conventional propagation of some species with medicinal value are: reduced endosperm, low germination rate and seedless varieties. The above mentioned reasons indicate the need for the production of artificial seeds as a technique which combines the advantages of clonal multiplication with those of seed propagation and storage. Objectives: The objective of the present investigation was to standardize artificial seed production technology taking shoot tip and nodal explants in Mentha arvensis and its in vitro regeneration Materials and Methods: Sodium alginate beads were produced by encapsulation of shoot tip and nodal segments of the plant M. arvensis. MS medium was used as basal medium with agar and sodium alginate was used as gelling agent accompanied by CaCl2 solution. Results: Different concentrations and combinations of BAP, Kin and NAA were used in alginate bead in MS basal medium. Among the different concentrations of phytohormone, highest 80% of shoot formation was observed in MS medium containing 2.0 mg/l BAP + 0.2 mg/l NAA from nodal segments of M. arvensis. Highest average number of shoot 9.87 ± 0.58 formation was obtained in the same medium but highest length of shoot 6.27 ± 0.29 cm was found in the medium having 1.0 mg/l BAP + 0.5 mg/l NAA. Conclusion: The present investigation clearly established and demonstrated the method of obtaining the artificial seed production in M. arvensis supported by different hormone concentrations DOI: http://dx.doi.org/10.3329/jbs.v20i0.17722 J. bio-sci.  20:  99-108, 2012


2017 ◽  
Vol 23 (1) ◽  
pp. 88 ◽  
Author(s):  
Paula Pinheiro Carvalho ◽  
Camila Aparecida Antoniazzi ◽  
Nayara Tayane Silva ◽  
Andréia Izabel Mikovski ◽  
Maurecilne Lemes Silva ◽  
...  

Passiflora miniata is a wild species native to the Southern Amazon, with ornamental potential due to the beauty of its flowers of intense red coloration. Reports in the literature about the species are still insipid. The aim of the present study was to induce the regeneration of P. miniata by the de novo organogenesis from mature zygotic embryos. The zygotic embryos were isolated and cultivated into the MS medium with the addition of 6-Benzyladenine (BA), Thidiazuron (TDZ) and Kinetin (KIN) growth regulators. The de novo regeneration from the zygotic embryos occurred directly and indirectly. A percentage of 80% of the explants cultivated in the presence of BA had direct organogenesis and 20% by the indirect way, with TDZ 60% were regenerated by the direct and 40% by the indirect way. Regarding the treatments with KIN, 58% of the explants had regeneration by direct and 42% by the indirect organogenesis. The development of shoot primordia initiated with the formation of organogenic structures that later differentiated into multi-shoots. The highest mean number of shoots (40.0 shoots per explants) was obtained on 0.75 mg L-1BA. Conversely, using 0.50 mg L-1 TDZ or KIN, the highest number of shoots were 7.2 and 3.6, respectively.


1970 ◽  
Vol 3 ◽  
Author(s):  
Mallapa Hanumanthu Niranjan ◽  
Mysore Shankar Sudarshana

The objective of this work was to study the histological events related to the regeneration process of a medicinal plant, Nymphoides cristatum (Roxb). Leaf explants were cultured on MS medium supplemented with 0.5 mgl-1 of 6-benzyl amino purine (BAP). About 90% of explants gave rise to shoots after 15 days of incubation. The histological studies showed that the regeneration originated directly from parenchymatous cells and direct organogenesis after 20 days of culture could be observed. Buds and roots were found completely differentiated after 40 days of culture and number of shoots per explants was 30. Micorshoots were rooted in hormone - free medium and the plants obtained grew in artificial pond under green house conditions. Key words: Leaf, Histology, in vitro regeneration, Nymphoides cristatum.  DOI: 10.3126/ijls.v3i0.2370


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sharone gladies E ◽  
Chithra Devi B. S

We can see Orchids come in a wide variety of shapes, sizes, colours, and textures far beyond the human mind’s imagination. They emerge from seeds in nature, but in the absence of suitable hosts, they do not germinate in sufficient numbers. This problem was solved by using the tissue culture technique for its germination. One of the successful method used for mass propogation of orchid plantlets is in vitro techniques. Therefore, an initial analysis was conducted in order to establish an appropriate procedure for mass multiplication of Arundina graminifolia. MS (Murashige and Skoog) medium was found to be suitable for the asymbiotic seed germination of Arundina graminifolia. Direct protocorm like bodies were induced by using combinations and individual supplement of MS medium with IAA (Indole-3-acetic acid), IBA (Indole-3- butyric acid), BAP (6-Benzylaminopurine) and KIN (Kinetin). Hormone-free MS basal medium was found suitable for the conversion of PLBs (protocorm-like bodies) into complete plantlets


HortScience ◽  
2018 ◽  
Vol 53 (1) ◽  
pp. 55-61 ◽  
Author(s):  
Mohsen Hesami ◽  
Mohammad Hosein Daneshvar

Ficus religiosa is an important industrial, medicinal, and ornamental plant, so in vitro regeneration is of high paramount in this valuable germplasm. Two efficient protocols were developed for indirect and direct shoot organogenesis through hypocotyl explants. In the first experiment, different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and indole butyric acid (IBA) (0.5, 1.0, and 1.5 mg·L−1) in combination with 6-benzyl amino purine (BAP) (ratio 10:1, respectively) were used for callus formation. Two types of callus were obtained from different concentrations of plant growth regulators (PGRs). Also, 2,4-D produced yellow-brownish and friable callus (Type I), whereas the green and compact callus (Type II) was achieved in IBA. The highest callus fresh weight (2.43 g) was observed in Murashige and Skoog (MS) medium containing 0.5 mg·L−1 2,4-D plus 0.05 mg·L−1 BAP. In the later experiments, various concentrations of thidiazuron (TDZ), 6-furfuryl amino purine (KN), and BAP (0.25, 0.5, 1.0, and 1.5 mg·L−1) in combination with IBA (ratio 10:1, respectively) were applied for shoot regeneration (direct and indirect organogenesis). In shoot regeneration from callus, the highest regeneration frequency (86.66%) and shoot number per callus (4.13) were achieved in MS medium supplemented with 1.5 mg·L−1 BAP plus 0.15 mg·L−1 IBA from type I calli. However, no regeneration was observed in type II calli. In direct shoot regeneration, the highest regeneration frequency (96.66%) and shoot number (6.26) were obtained in the medium mentioned previously. In root induction experiment, different concentrations of naphthalene acetic acid (NAA) and IBA alone or in combination were applied, and MS medium containing 2.0 mg·L−1 IBA along with 0.1 mg·L−1 NAA was the best hormonal balance for root induction. The rooted plantlets’ survival rate was more than 95% in the acclimatization stage. These results demonstrated that the direct regeneration method provides more shoot regeneration frequency and take a less time for shoot organogenesis than the indirect regeneration method. Based on our knowledge, this study is the first report of direct and indirect shoot organogenesis of F. religiosa via hypocotyl from in vitro–grown seedling.


2013 ◽  
Vol 22 (2) ◽  
pp. 179-185 ◽  
Author(s):  
SM Vidya ◽  
V Krishna ◽  
BK Manjunatha ◽  
MR Pradeepa

In vitro clonal propagation of Clerodendrum serratum L., a rare medicinal plant has been reported by using LM medium supplemented with different growth hormones. The maximum number of shoots with maximum length were obtained from stem derived callus on LM media fortified with 1.5 mg/l BAP and 0.3 mg/l NAA. Nodal explants showed direct organogenesis on LM media containing BAP (0.5 mg/l) alone. The regenerated shoots were successfully rooted with maximum frequency (100%) on half strength LM media supplemented with 0.5 mg/l NAA. The well rooted microshoots were successfully transferred to hardening and survival rate was 88%. DOI: http://dx.doi.org/10.3329/ptcb.v22i2.14208 Plant Tissue Cult. & Biotech. 22(2): 179-185, 2012 (December)


HortScience ◽  
2011 ◽  
Vol 46 (8) ◽  
pp. 1132-1135 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Rosario Julieta Baltazar-García ◽  
Victor Manuel Chávez-Avila

A protocol for in vitro propagation from protocorms of Oncidium tigrinum Llave & Lex., a threatened species distributed in Mexico and highly appreciated as an ornamental, was developed. Two different explants, entire protocorms and longitudinal halves of protocorms, were used. In addition, the effect of two different culture media, Murashige and Skoog (MS) and modified Knudson (KCm), supplemented with N6-benzyladenine (BA) (0, 0.5, 1, 2, 3, and 5 mg·L−1) and/or α-naphthaleneacetic acid (NAA) at 0, 0.1, and 0.5 mg·L−1 was investigated. Adventitious shoot formation by direct organogenesis was obtained in all treatments; in some cases, the formation of protocorm-like bodies was induced. Shoot formation was greater for entire protocorms; the best treatment was MS medium containing at BA 1 to 2 mg·L−1 in combination with at NAA 0.1 mg·L−1. The average height of shoots was three times greater in MS medium than in KCm medium. Subculturing individual shoots in MS medium without plant growth regulators, but with 1 g·L−1 activated charcoal, allowed further development and rooting. An ex vitro survival rate of almost 100% was achieved. This study represents a comprehensive application for propagation, conservation, and sustainable use of this valuable natural resource.


2021 ◽  
Vol 24 (1) ◽  
pp. 89-98
Author(s):  
SH Binto ◽  
JU Ahmed ◽  
TK Ghosh

Chinese fever vine (Paederia foetida L.), a valuable medicinal plant has been greatly utilized in therapeutic purposes throughout the world. Since conventional propagation techniques of P. foetida are very slow, inefficient and cannot cope with the increasing demand, in-vitro regeneration through tissue culture could be an alternative means of rapid propagation. Therefore, the efforts were made to develop a suitable protocol through direct organogenesis of P. foetida. After surface sterilization, the nodal explants were cultured in Murashigue and Skoog (MS) medium and MS medium supplemented with different concentrations and combinations of plant growth regulators. MS medium supplemented with 6-benzylaminopurine; BAP (2.0 mg L-1) produced the maximum number of shoots; 4.40 ± 0.98 and 5.40±1.12 after 15 and 30 days of culture respectively. The number of shoots gained by 15 days was found to be the highest; 1.20±0.80 at BAP (4.0 mg L-1) followed by 1.00±0.55 at BAP (2.0 mg L-1). Although the combination of BAP + Kinetin (2 mg L-1 +2 mg L-1) showed the highest shoot growth (3.40 ± 1.08 cm) by 15 days, sole application of BAP (2.0 mg L-1) or Kn (0.5, 1.0, 2.0 and 3.0 mg L-1) showed similar responses. BAP (2.0 mg L-1) showed the best responses for developing the highest number of leaves; 18.60 ± 2.42 and 29.20 ± 2.73 respectively after 15 and 30 days of culture. Similarly, development of the maximum number of leaves (10.60 ± 0.68) was reported by 15 days at BAP (2.0 mg L-1). Rooting was significantly induced in indole-3-acetic acid (IAA) supplemented to 1/2 strength MS medium as compared to control (only ½ strength MS medium). The best performance of rooting was observed by 0.5 mg L-1 IAA which produced average 4.33 roots per shoot after 21 days of culture. The regenerated plants showed similar morphology to the mother plants. Thus, a suitable protocol for successful multiplication of P. foetida in vitro was established using nodal explants. Ann. Bangladesh Agric. (2020) 24(1) : 88-98


2013 ◽  
Vol 18 (2) ◽  
pp. 25-33
Author(s):  
N Kamal ◽  
KM Nasuruddin ◽  
MS Haque ◽  
S Yasmin

An efficient and reproducible protocol for in vitro regeneration is required to achieve high frequency transformation from transformed calli. We report here high frequency plant regeneration from mature embryonic calli of two BINA rice cultivars Binasail and Binadhan-4. Embryonic calllus initiated on MS basal medium supplemented with 2 mg/l 2, 4-D. Several media with different combinations of growth regulators were tried. Maximum shoot regeneration frequency (63.33%) was observed in Binadhan-4 on MS medium supplemented with 5 mg/l Kinetin + 0.5 mg/l NAA. Maximum root regeneration frequency (70.00%) was observed in Binadhan-4 on MS medium supplemented with 6 mg/l Kinetin + 0.5 mg/l NAA. Well developed plantlets were hardened and transferred to the glasshouse.DOI: http://dx.doi.org/10.3329/pa.v18i2.17461 Progress. Agric. 18(2): 25 - 33, 2007


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