scholarly journals Influence of Additives on Enhanced In vitro Shoot Multiplication of Orthosiphon aristatus (Blume) Miq.

2013 ◽  
Vol 5 (3) ◽  
pp. 338-345 ◽  
Author(s):  
Swarna JAYAKUMAR ◽  
Ravindhran RAMALINGAM

Orthosiphon aristatus is a valuable medicinal plant and different parts of the plant are pharmaceutically used for the treatment of various diseases. The present study was designed to develop an efficient protocol for micropropagation of O. aristatus from nodal explants and to study the influence of additives on the enhancement of the number of shoots per explant. Among the different types of additives used, 10% coconut water and 30 mg/L glutamine added to Murashige and Skoog’s (MS) medium supplemented with 1.0 mg/L 6-benzyl amino purine (BAP) and 0.5 mg/L kinetin (KIN) was found to be most effective. Maximum number of shoots (44.07 ± 0.38) with 100% shooting response and shoot length of 7.47 ± 0.10 cm was recorded. In vitro rooting of the microshoots was achieved on half-strength MS medium containing 0.5 mg/L indole-3-butyric acid (IBA), producing an average of 30.27 ± 0.36 roots and 6.02 ± 0.20 cm root length. The rooted shoots were acclimatized with 100% survival rate on coco pith: soil (3:1) planting substrate and was successfully transferred to field conditions. The hardened plants exhibited homogeneity and no morphological variations were observed among the regenerants and the mother plants. Thus, the procedure described is a quick and reliable method which could be applied for efficient large-scale propagation, genetic transformation assays and secondary metabolite production.

Author(s):  
Girmay Mekonen ◽  
Meseret Chimdessa Egigu ◽  
Manikandan Muthsuwamy

Banana is a fruit crop which has high demand in Ethiopia, but its production is constrained by lack of disease free planting material with conventional propagation methods. For shoot initiation, shoot tip explants were cultured on MS medium supplemented with 0.5, 1.0, 1.5 and 2.0 mg/L BAP. Similarly, MS medium supplemented with BAP at 1.0, 1.5, 2.0 mg/L in combination with IBA at 0.25 and 0.50 mg/L were used for shoot multiplication. Half- strength MS medium augmented with IBA at 1.0, 2.0, 3.0 and 4.0 mg/l were used for root induction. MS medium without PGRs were used as controls. Finally, hardening of the in vitro derived plantlets was carried out in green house both in the primary and secondary acclimatization stages. Results showed that the highest shoot initiation percent (93.40%), highest mean number of shoots per explant (4.67) and lesser day for shoot induction (11.00) were observed in explant cultured on MS + 1.0 mg/L BAP. With shoot multiplication, highest shooting percent (92.60%), maximum number of shoots (7.67) and highest shoot length (5.27 cm) were recorded on MS + 1.5 mg/L BAP + 0.5 mg/L IBA. The highest rooting percent (93.40%), maximum root number per shoot (7.67) and highest root length (11.00 cm) were found on a half strength MS medium + 2.0 mg/L IBA. The survival rate of plantlets were 96.00% in coco peat substrate in primary acclimatization and 97.92% in forest soil, sand and manure substrates mixed at 3:2:1 ratio in secondary acclimatization. Overall, the result showed that the PGRs type, concentrations and combinations used are effective for mass propagation of banana variety studied in this experiment.


2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari

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.


2018 ◽  
Vol 30 (2) ◽  
pp. 283-294 ◽  
Author(s):  
Mani Manokari ◽  
Mahipal S. Shekhawat

Abstract The present study reports an efficient in vitro propagation system for Turnera ulmifolia using nodal segments as explants. Turnera ulmifolia (Passifloraceae) is an important garden plant with multipotent medicinal values. Effective shoot proliferation was achieved on agar gelled MS medium (Murashige and Skoog, 1962). The maximum number of shoots (8.3 ± 0.57) per initial explant was obtained on MS medium supplemented with 8.88 mM of 6-benzylaminopurine (BAP) and 0.54 mM of α-naphthalene acetic acid (NAA). The highest number of shoots (59.5 ± 2.10) proliferated on semi-solid MS medium (with agar) augmented with 2.22 mM of BAP and 2.32 mM of kinetin (Kin) along with 0.54 mM of NAA. Longer (4-5 cm) and healthy shoots were rooted (12.0 ± 0.10 roots per shoot) on half-strength MS medium fortified with 9.84 mM of indole-3 butyric acid (IBA). The in vitro regenerated plantlets were hardened in the greenhouse and transferred to the field. Significant developmental changes were observed in the foliar micromorphology of in vitro raised plantlets when these were transferred to the field. The stomatal index was gradually reduced (26.72 to 21.25) in the leaves from in vitro to field environments. But, vein-islets and veinlet terminations (13.4 and 7.6) were increased (39.7 and 18.4) respectively from in vitro to in vivo grown plants. Simple, unicellular, less frequent and underdeveloped trichomes were observed with the leaves of in vitro plants but fully developed trichomes recorded in the field transferred plants. The study could help in understanding the response and adaptation of tissue culture raised plantlets towards changed environmental conditions.


2021 ◽  
Vol 19 (1) ◽  
pp. 129-145
Author(s):  
Hoang Dac Khai ◽  
Nguyen Thi Nhu Mai ◽  
Hoang Le Lan Anh ◽  
Nguyen Nhu Minh Nguyet ◽  
Ho Viet Long ◽  
...  

Artichoke (Cynara scolymus L.), a medicinal plant with high economic value, contains high levels of phenolic compounds; especially cynarine, which plays an important role in preventing cancer, cardiovascular disease, osteoporosis, diabetes and neurodegeneration, etc. Currently, Artichoke micropropagation has achieved some success; however, the rooting efficiency and plantlet quality are still limited. In this study, improving the quality of Artichoke plantlet related to the shoot quality and suitable substrates in in vitro rooting stage was studied on “Violetta” Artichoke (VA) and “Green Globe” Artichoke (GA). The results showed that shoots (1.5 cm) cultured on MS medium supplemented 0.5 mg/L KIN were most suitable to shoot multiplication of VA with the number of shoots/explant (3.67 shoots), number of shoots ≥ 2 cm (3 shoots); while, 1.0 mg/L BA was suitable to shoot multiplication of GA (5.33 shoots; 5.00 shoots; respectively) after 4 weeks of culture. Besides, the in vitro rooting was improved using 8 g/L commercial agar for VA; meanwwhile, 3 g/L gelrite for GA. In addition, the nylon bag culture system (120 mm × 250 mm) has potential in plantlet production (15 plants/bag) and can be applied for large scale micropropagation. In addition, VA and GA plantlets derived from in vitro culture gave the good acclimatization, growth and development after 8, 12 and 20 weeks cultivating at the green house conditions.


2020 ◽  
Vol 100 (2) ◽  
pp. 202-208
Author(s):  
Mengting Wang ◽  
Guiliang Zhang ◽  
Peiyao Xin ◽  
Yun Liu ◽  
Bin Li ◽  
...  

Camellia fascicularis is an endangered evergreen ornamental plant with pale yellow flowers. An efficient and reproducible in vitro regeneration method is required for its large-scale propagation and germplasm conservation. In this study, one axillary bud per nodal stem was obtained from C. fascicularis cultured on Murashige & Skoog (MS) medium containing 0.1 mg L−1 indole-3-acetic acid (IAA) combined with 1.0 mg L−1 6-benzylaminopurine (BA). Axillary buds from the stem segments were transferred to modified woody plant medium (WPM) supplemented with 3.0 mg L−1 BA in combination with 0.3 mg L−1 IAA for multiplication, thereby resulting in a high shoot multiplication rate of 6.8. Multiple shoots were divided into nodal stems and shoot tips and were induced to root. The shoot tips were induced to root by culturing on one-half MS medium supplemented with 2.0 mg L−1 indole-3-butyric acid (IBA) in combination with 0.3 mg L−1 α-naphthalene acetic acid (NAA), which resulted in 76.0% rooting efficiency with 2.3 roots per shoot. The optimal hormone ratio for inducing rooting of nodal stems was 1.0 mg L−1 IBA in combination with 2.0 mg L−1 NAA, which resulted in 72.7% rooting efficiency with 1.7 roots per nodal stem. These two rooted plantlets were successfully acclimatized and established in a greenhouse.


2020 ◽  
Vol 51 (3) ◽  
pp. 976-981
Author(s):  
Al- Dabagh & Salih

 Technique of tissue culture for Chia (Salvia hispanica) micropropagation was achieved, this study investigated the impact of various concentrations of plant growth regulators on shoot multiplication and root induction with the Chia’s mature seed as a source explant. The highest percentage of shoot formation (80%), shoots number per explant(3.20) and shoot length(3.26 cm), were recorded on MS medium enriched with BAP(1.0 mgl-1) after eight weeks of seed culture. The optimal medium for the rhizogenesis was achieved on half strength MS medium fortified with 1.0 mgl-1 IBA after four weeks of culture, which had the highest rooting percentage (100%) with highest mean of roots number (5.6 roots per shoot) with (3.40 cm root length). The rooted plants were successfully adapted ex vitro with a survival rate of 85%.


2020 ◽  
Vol 19 (2) ◽  
pp. 67-73
Author(s):  
Majid Abdulhameed Ibrahim ◽  
Manal Zebari Sabty ◽  
Shaimaa Hussein Mussa

The study was conducted to mass micropropagation of big sage (Lantana camara L.) plant by shoot multiplication technique. The treatments 2.22 and 2.66 µmol·L–1 BA gave the highest significant increase in the percentage of response to shoot multiplication and number of shoots per explant compared to the other treatments as reached 96.70% and 100.00% and 4.33 and 6.00 shoots, respectively. The results showed that these two treatments did not differ significantly between them. While the 1.33 µmol·L–1 BA gave the lowest values in the percentage of response to shoot multiplication and number of shoots per explant were 80.00% and 2.00 shoots per explant, respectively. The MS medium supplemented with 4.30 or 5.37 µmol·L–1 NAA gave a high response to root formation, number of roots per shoot and root length. While the MS medium supplemented with 6.44 or 7.52 µmol·L–1 NAA gave low values in these characteristics. The MS medium with 2.22 or 2.66 µmol·L–1 concentration of BA or 7.52 µmol·L–1 concentration of NAA recorded the highest significant increase in the percentage of response to callus formation. While the MS medium supplemented with 1.33 µmol·L–1 BA or 4.30 µmol·L–1 NAA gave less response to the callus formation.


HortScience ◽  
2019 ◽  
Vol 54 (9) ◽  
pp. 1558-1564 ◽  
Author(s):  
Georgia Vlachou ◽  
Maria Papafotiou ◽  
Konstantinos F. Bertsouklis

Seed ecophysiology and micropropagation of Clinopodium nepeta, an aromatic Mediterranean plant with pharmaceutical and horticultural uses was investigated. The optimum germination temperature of seeds stored at room temperature for 0, 6, or 12 months was 15 to 20 °C (100% germination completed in 10 to14 days) and cardinal temperatures were defined at 10 and 30 °C (80% to 82% and 62% to 76% germination, respectively). Six or 12 months of storage did not seem to affect germination, although 12-month-old seeds germinated at higher percentage and completed germination earlier at 15 °C than at 20 °C. Concerning micropropagation, shoot multiplication at subcultures of both adult plant- and seedling-origin nodal explants was tested on Murashige and Skoog (MS) medium supplemented with various cytokinin types, i.e., zeatin (ZEA), 6-benzyladenine (BA), kinetin (KIN), and 6-γ-γ-(dimethylallylamino)-purine (2IP), at various concentrations from 0.0 to 8.0 mg·L−1. Both explant types presented a rather similar response during in vitro culture. Increasing concentration of all cytokinin types resulted in an increase in shoot number per responding explant (1.1–5.3) and in most cases a decrease in shoot length (0.6–3.4 cm). Increasing cytokinin concentration induced hyperhydricity to a number of shoots (0.1–6.5) per explant, mostly when ZEA and BA were used. Supplementing the MS medium with 8.0 mg·L−1 BA combined with 0.1 mg·L−1 1-naphthaleneacetic acid (NAA) led to almost elimination of hyperhydricity and very satisfactory shoot production (80%/88% explant response and 6.5/7.5 shoot number per responding explant for seedling- / adult-origin explants, respectively). Alternatively, increasing the agar concentration to 12.0 g·L−1 and supplementing the medium with 8.0 mg·L−1 BA only, resulted in the same effect on eliminating hyperhydricity, such as the addition of NAA, and in the best shoot multiplication response achieved in this study (100% explant response, 9.4/9.9 shoots per explant for seedling-/adult-origin explants, respectively). Microshoots rooted abundantly (92% to 100%) on half-strength MS medium, either Hf or supplemented with 0.5 mg·L−1 to 4.0 mg·L−1 indole-3-butyric acid (IBA). The addition of IBA to the rooting medium, regardless of its concentration, affected only the root length by increasing it 2- to 3-fold. Microshoot clusters produced on multiplication media rooted at 96% when cultured on Hf half-strength MS medium. Rooted microshoots and shoot clusters survived at 80% to 100%, respectively, after ex vitro acclimatization in peat:perlite 1:1 (v/v).


Author(s):  
Sanjay Kumar Madkami ◽  
Arpita Moharana ◽  
Durga Prasad Barik

Background: The presence of L-dopa coupled with rich protein and amino acid marked Mucuna pruriens var. utilis as an important under-utilised legume. Therefore, it is useful to develop a method for large-scale multiplication for commercial production. Method: Tissue culture technology is successfully utilized in propagation of plants with poor and uncertain response to conventional propagation. Murashige and Skoog’s (MS) medium without any Plant Growth Regulators (PGRs) was used for seed germination and with PGRs for shoot and root multiplication.Result: Highest 95% seed germination was found in fresh seeds at 7-8 days of culture. Shoot multiplication percentage was found to be 100% with highest c.a. 21.1 shoots with an average 4.8 cm shoot length on MS + BAP 1.5 mg L-1 per 10 days old cotyledonary node explant. A total c.a. 144 shoots were harvested after 3rd harvest of the mother cotyledonary node with two whole cotyledons at day 70. Rooting was best induced in in vitro derived shoots on MS medium without any PGRs and plantlets were acclimatized in sand and soil (1:1) and established in pot with garden soil. 


Scientifica ◽  
2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari ◽  
C. P. Ravindran

A tissue culture protocol was developed for an important medicinal plantRungia pectinataL. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2solution. Murashige and Skoog (MS) medium was used to establish the cultures ofR. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L−16-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L−1each of BAP and kinetin (Kin) + 0.1 mg L−1indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L−1indole-3 butyric acid (IBA).In vitroflowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots underin vitroconditions.


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