scholarly journals COMPARISON OF TWO METHODS FOR IN VITRO PROPAGATION OF RAUWOLFIA SERPENTINA FROM NODAL EXPLANTS

2007 ◽  
Vol 44 (07) ◽  
pp. 514-519 ◽  
Author(s):  
Ved Prakash Pandey ◽  
Jose Kudakasseril ◽  
Elizabeth Cherian ◽  
George Patani ◽  

Two different methods of in vitro multiplication of Rauwolfia serpentina from nodal explants were compared viz. multiplication via callus morphogenesis and that via shoot proliferation from axillary buds. The second method was found to be far better. The optimum shoot proliferation occurred on Murashige and Skoog (MS) medium supplemented with 1 mg/L naphthalene acetic acid (NAA) and 2 mg/L of benzyl aminopurine (BAP). The best rooting of shoots occurred on MS medium containing 4% sucrose and 1 mg/L of NAA. Solid and liquid MS media were found to be similar in supporting shoot proliferation. The plants produced were successfully hardened and established in soil. An easy, reliable and reproducible protocol was developed for in vitro micropropagation of Rauwolfia serpentina from nodal explants.

2020 ◽  
Author(s):  
Nurşen Çördük ◽  
Cüneyt Aki

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.


Biologia ◽  
2014 ◽  
Vol 69 (5) ◽  
Author(s):  
Ziba Bakhtiar ◽  
Mohammad Mirjalili ◽  
Ali Sonboli ◽  
Mahdi Farimani ◽  
Mahdi Ayyari

AbstractThymus persicus (Ronniger ex Rech. f.) Jalas is a valuable and endangered natural source of antitumor pentacyclic triterpenoids, i.e., betulinic acid, oleanolic acid and ursolic acid, which grows in northwest Iran. As the plant has a low propagation rate in nature, a suitable method for in vitro-propagation is needed. With the aim of identifying a suitable system for regenerating T. persicus via direct organogenesis, Murashige & Skoog (MS) medium supplemented with different plant growth regulators (PGRs) was tested. In vitro-grown shoot tips were exposed to the cytokinins 6-benzylaminopurine (BAP), kinetin (KN), and thidiazuron (TDZ), alone or in combination with the auxins 1-naphthalene-acetic acid (NAA), 2,4-dichlorophenoxyacetic (2,4-D), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). The highest shoot formation (7.1 ± 0.9) was obtained with a medium fortified with 8.9 μM BAP plus 2.7 μM NAA. Regenerated shoots were easily rooted on the different tested media, with the most abundant (16.6 ± 1.4) and strongest roots obtained on half-strength MS medium containing 2.5 μM IBA. The rooted plantlets were successfully acclimatized (76.6%) in a greenhouse before transference to natural conditions. Homogeneity and phytochemical productivity of the in vitro regenerated plantlets were confirmed by random amplified polymorphic DNA (RAPD) profiles and high performance liquid chromatography (HPLC), respectively.


2016 ◽  
Vol 4 (2) ◽  
pp. 155-160
Author(s):  
Lila Bahadur Magar ◽  
Nisha Shrestha ◽  
Saraswoti Khadka ◽  
Jay Raj Joshi ◽  
Jibaraj Acharya ◽  
...  

Paulownia tomentosa Steud is a fast growing multipurpose tree. In vitro propagation using nodal explants of Paulownia tomentosa was performed by manipulating amount of cytokinin and auxin in culture media. Shoot bud proliferation from explants were assessed in Murashige and Skoog (MS) medium supplemented with various amounts of hormones such as a) 0.5-2.5 mg/l benzylaminopurine (BAP) and 0.1 mg/l naphthalene acetic acid (NAA), b) 0.5-2.5 mg/l BAP and 0.5-2.5 mg/l kinetin (KN) and c) 0.5- 2.5 mg/l BAP and 0.1 mg/l indole-3-acetic acid (IAA). In the present study, we found that hormone combination of BAP and NAA gave optimum growth results. MS medium enriched with 2.0 mg/l BAP and 0.1 mg/l IAA resulted a similar outcome but took 3-4 weeks with respect to the same medium enriched with 1.0 mg/ml BAP and 0.1 mg/ml NAA, which showed response within 2-3 weeks. Shoot length of 2.5-3.5 cm with 3-4 nodes and 8-12 leaves were obtained on MS medium supplemented with 1.0 mg/l BAP and 0.1 mg/l NAA. The acclimatization of explants was done in a polyhouse at 20±5oC for 2-6 weeks. Rooting has been induced in nonsterile sand. Rooted plants were transferred to plastic bags containing mixture of soil, sand and compost in the ratio of 1:1:1.Besides aforementioned issues, there are several other challenges associated with in vitro propagation of P. tomentosa. The plants were established (90%) on MS medium enriched with BAP and NAA and adapted ex vitro with surviving up to 80%. People received an opportunity with this plant because it grows fast and can generate income in 10 years in comparison with others, but at the same time people also have been facing the challenges for plantation of P. tomentosa as government of Nepal does not formulate necessary national policies, legislations and regulatory frameworks in its favor. Thus, system should be developed to set royalty rate of P. tomentosa recognizing its lifetime value.Int J Appl Sci Biotechnol, Vol 4(2): 155-160


2015 ◽  
Vol 22 (1) ◽  
pp. 33-36
Author(s):  
A. Benniamin ◽  
G. Jothi ◽  
M. Sundari

Protocols of axillary bud multiplication were established for Dodonaea viscosa (Sapindaceae). Murashig & Skoog’s medium with 0.2 mg/l BAP with 0.01 Naphthalene acetic acid induced high rate of shoot induction and an average of five shoots per node. Subsequent culture enhanced the number of shoots. Callus initiated from the basal cut end explants differentiated in to more than 10 shoots on MS Medium with 0.4mg/l Benzylaminopurine and 0.02mg/l Indole Butric Acid. Eighty per cent of the rooted shoots survived when transferred to greenhouse and subsequently to the field.


2020 ◽  
Vol 44 (4) ◽  
pp. 641-648
Author(s):  
Bandita Deo ◽  
Bikram Keshari ◽  
Bikram Pradhan

The present experiment was conducted to optimize protocols for in vitro propagation of banana (Musa sp.) cv. ‘Patakpura’ (AAB), supplemented with different growth regulators. Shoot tips obtained from sword suckers were cultured aseptically on MS medium supplemented with different concentrations of cytokinins like 6-Benzylaminopurine (BAP) and Kinetin (KN) for multiplication of shootsand auxins such as indole acetic acid (IAA) and naphthalene acetic acid (NAA) for induction of roots. The best result from the initial culture was obtained from MS medium supplimented with 4 mg/l BAP + 0.5 mg/l IAA. The highest shoot fresh weight, shoot length and number of shoots per explant were recorded from MS medium supplemented with 4 mg/l BAP + 0.5 mg/l IAA + 0.25 mg/l NAA. Therefore, the MS medium supplemented with 4 mg/l BAP + 0.5 mg/l IAA + 0.25 mg/l NAA was found to be most effective and productive combination for shoot multiplication and proliferation of the culture in vitro. IAA at a concentration of 1 mg/l was found to be most suitable for rooting of the shoots. Bangladesh J. Agril. Res. 44(4): 641-648, December 2019


HortScience ◽  
2005 ◽  
Vol 40 (6) ◽  
pp. 1832-1837 ◽  
Author(s):  
Donita L. Bryan ◽  
Michael A. Arnold ◽  
R. Daniel Lineberger ◽  
W. Todd Watson

Three spineless phenotypes of Acacia wrightii G. Bentham ex A. Gray were identified with aesthetic landscape potential. Experiments in seed, cutting, grafting, and tissue culture propagation were undertaken to perpetuate this desired spineless phenotype. Germination percentages for mechanically scarified seeds ranged from 33% to 94%, however yield of spineless seedlings was low (0% to 34%). Sulfuric acid scarification for 10, 20, 30, or 60 minutes hastened and unified germination compared to nontreated seeds by 7 to 8 days. Vegetative propagation was successful for softwood cuttings. Rooting measures increased with auxin (2:1 indole butyric acid to naphthalene acetic acid) concentrations from 0 to 15000 mg·L–1, with maximum rooting percentage (70%), root number (9.2), and root length (12.4 cm) per softwood cutting at 15000 mg·L–1 auxin 8 weeks after treatment. Rooting was not successful for semi-hardwood or hardwood cuttings. Whip-and-tongue or T-bud grafting was not successful. Tissue culture of shoots from in vitro germinated seedlings indicated that shoot proliferation was greatest in Murashige and Skoog (MS) medium with 15 μm zeatin. The number of shoots that rooted in vitro increased with increasing concentrations of indole-3-butyric acid from 0 to 25 μm.


HortScience ◽  
2016 ◽  
Vol 51 (7) ◽  
pp. 905-909 ◽  
Author(s):  
Jane Kahia ◽  
Peter Njenga ◽  
Margaret Kirika

The effect of phytohormones on breaking of dormancy of axillary buds in anchote and their subsequent shoot proliferation were examined. Anchote is an annual trailing endemic plant with high calcium content grown for its edible tuberous roots in Ethiopia. Nodal explants harvested from the greenhouse were sterilized using various concentrations of a commercial bleach (JIK) which contains 3.85% sodium hypochlorite (NaOCl) and time duration. The highest (85%) clean explants were obtained when 5% JIK was used for 15 minutes. The explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of benzylaminopurine (BAP), kinetin, and thidiazuron (TDZ). The highest frequency of microshoot induction (84%) and mean number of microshoots (3.4) were recorded from explants cultured on medium supplemented with TDZ 0.025 µm. Hyperhydrated shoots were observed on media supplemented with high concentrations of BAP and kinetin but interestingly not on TDZ media. Induction of roots was highest (86%; 4.6 roots per shoot) when shoots were transferred to half strength MS medium containing 0.5 μm α-naphthalene acetic acid (NAA) after 12 days. A survival rate of 83% was recorded in the greenhouse and the plantlets appeared to be morphologically normal. This is the first report on use of TDZ for in vitro propagation of anchote.


Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 246
Author(s):  
Shipra Rani Jha ◽  
Ruphi Naz ◽  
Ambreen Asif ◽  
Mohammad K. Okla ◽  
Walid Soufan ◽  
...  

An efficient protocol of plant regeneration through indirect organogenesis in Viola serpens was developed in the present study. Culture of leaf explants on MS (Murashige and Skoog) medium supplemented with 2.0 mg/L 6-benzyladenine and 0.13 mg/L 2,4-dichloro phenoxy acetic acid. Adventitious shoot formation was observed when calli were transferred on to MS medium containing 0.5 mg/L α-naphthalene acetic acid and 2.25 mg/L kinetin, which showed the maximum 86% shoot regeneration frequency. The highest root frequency (80.92%) with the 5.6 roots per explant and 1.87 cm root length was observed on MS medium supplemented with 2 mg/L indole-3-butyric acid. The plantlets were transferred to the mixture of sand, coffee husk and soil in the ratio of 1:2:1 in a pot, and placed under 80% shade net for one month. It was then transferred to 30% shade net for another one month, prior to transplantation in the field. These plantlets successfully acclimatized under field conditions. A Sequence Characterized Amplified Region (SCAR) marker was also developed using a 1135 bp amplicon that was obtained from RAPD (Random Amplification of Polymorphic DNA) analysis of six accessions of V. serpens. Testing of several market samples of V. serpens using the SCAR marker revealed successful identification of the genuine samples of V. serpens. This study, therefore, provides a proficient in vitro propagation protocol of V. serpens using leaf explants and a SCAR marker for the authentic identification of V. serpens. This study will be helpful for conservation of authentic V. serpens.


2018 ◽  
Vol 16 (1) ◽  
pp. 27-30
Author(s):  
Farhana Hoque ◽  
Mahbub Robbani ◽  
Md Fakhrul Hasan ◽  
Jahanara Parvin

An experiment was conducted at the Plant Biotechnology Laboratory, Department of Horticulture, Patuakhali Science and Technology University during the period from August 2016 to April 2017 to standardize the protocol for in vitro propagation of banana. The experiment was laid out in completely randomized design with four replications. Three to four months aged field grown rhizome attached shoots were used as explants and cultured on MS medium with different concentrations and combinations of BAP (6-benzylamino purine), BAP + KIN (Kinetin) + NAA (Naphthalene Acetic Acid) and IBA (Indole-3- Butyric Acid) + IAA (Indole-3- Acetic Acid) to observe their efficacy on single shoot initiation, shoot multiplication and root formation respectively. Minimum number of days required for shoot initiation (9.07) with highest shoot initiation percentage (91.14) and the longest shoot (2.23 cm) was found in MS medium supplemented with 5.0 mg/L BAP. On the other hand, highest shoot multiplication percentage (80.99) with maximum number of shoots per explant (4.47), the highest length of shoots (4.17 cm) and maximum number of leaves (4.04)was observed in MS medium supplemented with 4.0 mg/L BAP + 2.0 mg/L KIN + 2.0 mg/L NAA. In case of root regeneration, the best results on days required for root initiation (9.00), the highest root initiation percentage (85.05), maximum number of roots per plantlet (5.83) and the highest length of roots (4.17 cm) was obtained in MS medium supplemented with1.5 mg/L IBA + 0.5 mg/L IAA. After 5-7 days of hardening in room temperature, established plantlets were ready for plantingJ. Bangladesh Agril. Univ. 16(1): 27-30, April 2018


HortScience ◽  
1994 ◽  
Vol 29 (3) ◽  
pp. 214-216 ◽  
Author(s):  
S. Singh ◽  
B.K. Ray ◽  
S. Bhattacharyya ◽  
P.C. Deka

Multiple shoots were obtained from shoot tips (2 to 3 mm) derived from mature plants (5 to 6 years old) of Citrus reticulata Blanco cv. Khasi mandarin and C. limon Burm.f. cv. Assam lemon when cultured on Murashige and Skoog (MS) medium, supplemented with (mg·liter-1) 1.0 BAP, 0.5 kinetin, and 0.5 NAA. Root induction was observed when 7-week-old single shoots (≈ 2 cm long) of both Citrus species were cultured on MS medium supplemented with (mg·liter-1) 0.25 BAP, 0.5 NAA, and 0.5 IBA. These plantlets were successfully established in the soil. Chemical names used: naphthalene acetic acid (NAA), indole 3-butyric acid (IBA), and benzylamino purine (BAP).


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