scholarly journals EFFECT OF SUCROSE AND MEDIA STRENGTH ON IN VITRO MULTIPLICATION IN SWERTIA CHIRATA BUCH.-HAM EX WALL: AN ENDANGERED MEDICINAL HERB

2018 ◽  
Vol 10 (11) ◽  
pp. 40
Author(s):  
Manu Pant ◽  
Prabha Bisht ◽  
Manju P Gusain
Keyword(s):  
Shoot Multiplication ◽  
Axillary Shoot ◽  
Ms Medium ◽  
Carbohydrate Source ◽  
Windows Nt ◽  
Swertia Chirata ◽  
Shoot Number ◽  
Endangered Medicinal Herb ◽  
Experimental Findings

Objective: The present study was performed to investigate the role of varying concentrations of carbohydrate source and strengths of nutrient medium in growth and development of in vitro shoots of Swertia chirata-an endangered medicinal plant.Methods: MS medium supplemented with 6-Benzylaminopurine (4.44 µM), Indole-3 acetic acid (2.85 µM) and Adenine sulphate (271.45 µM) was used to test the efficiency of of sucrose at concentrations of 1-5% and of media strength varying from full to one-fourth. The data were analysed using analysis of variance (ANOVA) of Completely Randomized Design (CRD) in GenStat 5 Edition 3.2 for PC/Windows NT (Copyright 1995, Lawes Agricultural Trust (Rothamsted Experimental Station))Results: Observations on axillary shoot multiplication indicated that sucrose at a concentration of 3% and MS medium in its full strength proved to be most optimal for in vitro culture multiplication. On this medium combination mean number of 11.80 shoots (after 4 w) and 18.50 shoots (after 8 w) could be obtained On sucrose free medium the shoots exhibited necrosis while at lower concentrations of 1-2% sucrose, the shoots developed were thin and unsuitable for further growth in vitro. At higher levels of sucrose in the medium, the shoots became thick and stunted. Similarly, reduction in medium strength resulted in a decline in shoot number and shoot length to an average of 6.50 shoots (1.33 cm mean length) on half strength medium and 5.60 shoots (0.88 cm mean length) on one-fourth strength; as observed after 4 w.Conclusion: The experimental findings suggest that any decline from the standard had a significant effect on the number, size and overall health of shoots developed in vitro. The conditions so standardized augment the production of healthy shoots that shall aid in subsequent rooting and survival after transplantation of tissue-culture raised plantlets.

scholarly journals Micropropagation of Clerodendrum phlomidis L.F.

2016 ◽  
Vol 24 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Mafatlal M. Kher ◽  
Deepak Soner ◽  
Neha Srivastava ◽  
Murugan Nataraj ◽  
Jaime A. Teixeira da Silva
Keyword(s):  
Callus Formation ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Explants ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoot Proliferation ◽  
Clerodendrum Phlomidis

Abstract Clerodendrum phlomidis L. f. is an important medicinal plant of the Lamiaceae family, particularly its roots, which are used for various therapeutic purposes in a pulverized form. The objective of this study was to develop a standard protocol for axillary shoot proliferation and rooting of C. phlomidis for its propagation and conservation. Nodal explants were inoculated on Murashige and Skoog (MS) medium that was supplemented with one of six cytokinins: 6-benzyladenine, kinetin, thidiazuron, N6-(2-isopentenyl) adenine (2iP), trans-zeatin (Zea) and meta-topolin. Callus induction, which was prolific at all concentrations, formed at the base of nodal explants and hindered shoot multiplication and elongation. To avoid or reduce callus formation with the objective of increasing shoot formation, the same six cytokinins were combined with 4 μM 2,3,5-tri-iodobenzoic acid (TIBA) alone or in combination with 270 μM adenine sulphate (AdS). Nodal explants that were cultured on the medium supplemented with 9.12 μM Zea, 4 μM TIBA and 270 μM AdS produced significantly more and longer shoots than on medium without TIBA and AdS. Half-strength MS medium supplemented with 8.05 μM α-naphthaleneacetic acid was the best medium for root formation. Most (75%) in vitro rooted plantlets were successfully acclimatized under natural conditions.

scholarly journals Studies on Seed Germination and Micropropagation of Clinopodium nepeta: A Medicinal and Aromatic Plant

HortScience ◽  
2019 ◽  
Vol 54 (9) ◽  
pp. 1558-1564 ◽  
Author(s):  
Georgia Vlachou ◽  
Maria Papafotiou ◽  
Konstantinos F. Bertsouklis
Keyword(s):  
Shoot Multiplication ◽  
Adult Plant ◽  
Naphthaleneacetic Acid ◽  
Similar Response ◽  
Ms Medium ◽  
Rooting Medium ◽  
Shoot Number ◽  
Half Strength ◽  
Mediterranean Plant

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).

scholarly journals In Vitro Regeneration of Grass Pea (Lathyrus sativus L.) from Cotyledonary Node Explants

2017 ◽  
Vol 5 (2) ◽  
pp. 57 ◽  
Author(s):  
Diriba Tesfaye ◽  
Kassahun Bantte ◽  
Tewodros Tadesse
Keyword(s):  
In Vitro Regeneration ◽  
Shoot Multiplication ◽  
Full Potential ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Grass Pea ◽  
In Vitro Techniques ◽  
Shoot Initiation ◽  
Shoot Number

Full potential of grass pea has not been utilized because of the presence of the neurotoxin amino acid β-N-oxalyl-L-αβ -diaminopropionic acid (ODAP/BOAA). Conventional breeding and other approaches have not been successful in reducing the toxin. Integration of in vitro techniques can contribute significantly to meet the challenge. Therefore, this study was carried out to evaluate the in vitro regeneration capacity of grass pea genotypes. Shoot initiation, multiplication and rooting of IVAT-LS-690 were conducted using completely randomized design with five replications. Genotypes were treated with BAP and NAA for shoot initiation while BAP and Kn Combination were used for multiplication. Different concentrations of IBA and IAA were used for rooting. Shoot proliferation percentage was the highest (100%) for IVAT-LS-690,on Murashige and Skoog (MS) medium augmented with 2.0 mg/l BAP +0.1 mg/lNAA.For in vitro shoot multiplication, best results were obtained on concentrations of 3mg/l BAP+1mg/l Kn with maximum shoot number per explants (11.5). High number of roots per shoot (6) and percent of rooted shoot (86.66%) were obtained from ½ MS medium supplemented with 0.5 mg/l IBA. This study inferred that both genotype and BAP levels play a crucial role for shoot regeneration capacity and the optimum hormonal combination for grass pea is genotype specific.

scholarly journals Study on in vitro propagation of Giao co lam (Gynostemma pentaphyllium Thunb.)

2018 ◽  
Vol 17 (05) ◽  
pp. 84-92
Author(s):  
Trinh L. D. Ho
Keyword(s):  
Leaf Explants ◽  
Shoot Multiplication ◽  
Young Leaf ◽  
Ms Medium ◽  
Nodal Explant ◽  
Coconut Fiber ◽  
Average Percentage ◽  
Endangered Medicinal Herb ◽  
Multiplication Score

Giao co lam (Gynostemma pentaphyllium Thunb.) is a traditional medicine plant and endangered species in Vietnam. The research was carried out to establish the plant propagation for the purpose of conserving and exploiting this endangered medicinal herb. The young leaf and nodes of Giao co lam in vitro were used as explants in the study to evaluate the factors influencing the multiplication. Young leaf explants were excised and cultured in MS medium with TDZ from 0.1 to 1 mg/L. After 10 weeks of culture, new shoots came out from their explants and the MS medium containing TDZ 0,7 mg/L gave the highest shoots (12.89 shoots/explant) with the average percentage of 74.67%. When nodal explants were cultured on MS supplemented with BA at a concentration of 0.3 to 1.5 mg/L and IBA 0.5 mg/L. After 6 weeks of culture, explants on MS medium supplemented with BA 1 mg/L and IBA 0,5 mg/L gave the highest shoots (7.39 shoots/explant) and their average percentage was 83.33%. In comparison to the nodal explant medium in combination with BA (0.5 to 3 mg/L) and NAA 0.2 mg/L for 4 weeks of culture, the best rapid shoot multiplication score was 3.67 times with MS + BA 1.5 mg/L + NAA 0.2 mg/L as compared to MS + BA 1.0 mg/L + IBA 0.5 mg/L. Suitable medium for rooting was MS + 0.5 mg/L IBA with root shoots at 97.33%, average roots at 5.29 roots/shoot after 4 weeks. On organic substrat, 70% coconut fiber and 30% composted cow manure gave the highest survival rate of 91.33%. The plants grew and developed well during the nursery stage.

scholarly journals 586 PB 123 AXILLARY SHOOT REGENERATION IN CHINKAPIN OAK

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 515g-515
Author(s):  
Sudeep Vyapari ◽  
Houchang Khatamian
Keyword(s):  
Shoot Regeneration ◽  
Shoot Multiplication ◽  
Shoot Tip ◽  
Axillary Shoot ◽  
Ms Medium ◽  
Pulse Treatment ◽  
Axillary Shoots ◽  
Shoot Tip Explants

Surface disinfested nodal and shoot-tip sections of chinkapin oak (Quercus muehlenbergii Engelm.), obtained from adult or juvenile source, when cultured on WFM supplemented with BA or kinetin (1.0 -5.0 mg l-1) produced greater number of axillary shoots per explant and shoot lengths than MS medium. Nodal and shoot-tip explants cultured in WPM containing cytokinins, BA or kinetin (0.1 - 5.0 mg l“) resulted in greater number of axillary shoots than media containing auxins, 2,4-D or NAA (1.0 - 5.0 mg l-1). In vitro grown shoot explants cultured in WFM shoot multiplication medium containing thidiazuron did not produce axillary shoots. Microshoots when cultured in WFM plus NAA or IBA (0.1 -2.0 mg l-1), or subjected to IBA (0.5 mg l-1) pulse treatment (0, 5, 10 or 15 min.) did not root.

scholarly journals In Vitro Propagation of Alexandrian Laurel (Danae racemosa L. Moench), a Valuable Ornamental Plant

HortScience ◽  
2013 ◽  
Vol 48 (10) ◽  
pp. 1301-1303
Author(s):  
Xiuli Shen ◽  
Guochen Yang ◽  
Zhongge (Cindy) Lu
Keyword(s):  
Morphological Variation ◽  
In Vitro Propagation ◽  
Shoot Multiplication ◽  
Ornamental Plant ◽  
Efficient Production ◽  
Ms Medium ◽  
Culture Initiation ◽  
Shoot Number ◽  
Alternative Means

To overcome the limitations of traditional propagation, this research was initiated to develop an alternative means for efficient production of Alexandrian laurel (Danae racemosa L. Moench). An in vitro propagation protocol has been developed for Danae racemosa L. Moench using seeds as a source of material for culture initiation. Seedlings were produced after seeds were cultured for 3 month on MS (Murashige and Skoog, 1962) medium. Shoot multiplication occurred on MS medium with or without 6-benzylaminopurine (BAP) with 100% multiplication percentage. However, shoot number was significantly increased from an average of 2.8 to more than six with the addition of 5 or 25 μM BAP. Among two indole-3-butyric acid (IBA) treatments tested for rooting of seedlings, incorporation of 5 μM IBA in MS medium significantly increased rooting percentage to 86.4% compared with 71.2% without IBA. The greatest number of roots (three) was produced by 5-minute IBA pulse. However, both IBA treatments significantly reduced root length. The longest root (12.8 mm) was observed on MS medium without any IBA treatment and the shortest (6.1 mm) was produced by IBA pulse. In vitro-propagated plantlets grew well after transfer to a substrate of peat and pine bark (1:1) in the greenhouse. No morphological variation was observed.

scholarly journals Optimizing micropropagation conditions for a recalcitrant ninebark (Physocarpus opulifolius L. maxim.) cultivar

2021 ◽  
Vol 57 (2) ◽  
pp. 281-295 ◽  
Author(s):  
K. Jagiełło-Kubiec ◽  
K. Nowakowska ◽  
A. Ilczuk ◽  
A. J. Łukaszewska
Keyword(s):  
Basal Medium ◽  
Cost Effective ◽  
Ms Medium ◽  
Carbohydrate Source ◽  
Shoot Production ◽  
Shoot Number ◽  
Plant Acclimation ◽  
Physocarpus Opulifolius ◽  
Half Strength

AbstractNinebark is a very popular ornamental shrub. Micropropagation is an efficient method for mass production of uniform plant material. This study was designed to develop and optimize conditions at all phases of ninebark micropropagation. For the multiplication stage, the Murashige and Skoog (MS) medium at full concentration and pH 5.8 was chosen as the basal medium. Sorbitol proved a more effective carbohydrate source than fructose, with no adverse effects on shoot vitrification or the medium itself. The best shoot production, both in number and length, was on the medium enriched with 2 and 3 mg·L−1 zeatin. High numbers of shoots were also obtained in treatments with 1 mg·L−1 6-benzyladenine (BA) or 2 mg·L−1 meta-Topolin (mT) in the basal medium. BA was the most cost-effective cytokinin. There was a positive effect of the gibberellic acid on proliferation: the highest shoot number per explant was produced in the presence of 1 mg·L−1 GA3. No effect of the culture age (up to 20 subcultures) on the percentage of regenerating explants was evident, and the highest numbers of shoots were obtained between passages 10 and 17. For rooting, the MS medium at half strength was used. The best rooting was at 1 mg·L−1 IBA. Spraying the in vitro rooted cuttings with abscisic acid (ABA) favored plant acclimation to the ex vitro conditions. Exvitro rooting, including the treatments with IBA and ABA, shortened the production time by approximately one third.

scholarly journals Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3229
Author(s):  
Mat Yunus Najhah ◽  
Hawa Z. E. Jaafar ◽  
Jaafar Juju Nakasha ◽  
Mansor Hakiman
Keyword(s):  
Callus Induction ◽  
Antioxidant Activities ◽  
Shoot Multiplication ◽  
Wild Plants ◽  
Wild Plant ◽  
Total Phenolic ◽  
Nodal Segment ◽  
Ms Medium ◽  
In Vitro Plantlets

This study aims to investigate whether the in vitro-cultured L. pumila var. alata has higher antioxidant activity than its wild plant. An 8-week-old L. pumila var. alata nodal segment and leaf explants were cultured onto Murashige and Skoog (MS) medium supplemented with various cytokinins (zeatin, kinetin, and 6-benzylaminopurine (BAP)) for shoot multiplication and auxins (2,4-dichlorophenoxyacetic acid (2,4-D) and picloram) for callus induction, respectively. The results showed that 2 mg/L zeatin produced the optimal results for shoot and leaf development, and 0.5 mg/L 2,4-D produced the highest callus induction results (60%). After this, 0.5 mg/L 2,4-D was combined with 0.25 mg/L cytokinins and supplemented to the MS medium. The optimal results for callus induction (100%) with yellowish to greenish and compact texture were obtained using 0.5 mg/L 2,4-D combined with 0.25 mg/L zeatin. Leaves obtained from in vitro plantlets and wild plants as well as callus were extracted and analyzed for their antioxidant activities (DPPH and FRAP methods) and polyphenolic properties (total flavonoid and total phenolic content). When compared with leaf extracts of in vitro plantlets and wild plants of L. pumila var. alata, the callus extract displayed significantly higher antioxidant activities and total phenolic and flavonoid content. Hence, callus culture potentially can be adapted for antioxidant and polyphenolic production to satisfy pharmaceutical and nutraceutical needs while conserving wild L. pumila var. alata.

scholarly journals Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile

2019 ◽  
Vol 11 (2) ◽  
Author(s):  
POPY HARTATIE HARDJO ◽  
DANNY PUTRA SENTOSA SUSANTO ◽  
WINA DIAN SAVITRI ◽  
MARIA GORETTI MARIANTI PURWANTO
Keyword(s):  
Growth Index ◽  
Shoot Multiplication ◽  
High Price ◽  
Ms Medium ◽  
Ex Vitro ◽  
Average Growth ◽  
In Vitro Multiplication ◽  
Patchouli Alcohol ◽  
Pogostemon Cablin

Abstract. Hardjo PH, Susanto DPS, Savitri WD, Purwanto MGM. 2019. Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile. Nusantara Bioscience 11: 123-127. Pogostemon cablin Benth. is a plant producing patchouli oil, which mostly consists of patchouli alcohol compound. Patchouli oil has great potentials in the world market because of its stability and high price. In this study, in vitro multiplication of Sidikalang variety of Acehnese patchouli shoots was done on solid and liquid Murashige & Skoog (MS) medium. This study aimed to determine the effect of cytokinins in various combinations of shoot multiplication and to compare the patchouli oil yield of in vitro and ex vitro culture. In vitro multiplication of Acehnese patchouli shoots by using solid MS medium with addition of 0.2 ppm benzyl aminopurine (BAP) and 0.2 ppm Kinetin resulted in shoot explants with an average growth index of 82.198 ± 0.690. Patchouli oil extraction was done on 7 weeks old in vitro shoot explants cultured on solid MS medium + 0.2 ppm BAP + 0.2 ppm Kinetin using water distillation method. In vitro shoots yielded 2.5% patchouli oil and contained ± 35% patchouli alcohol compound, whereas ex vitro shoots produced 4% patchouli oil and contained ± 25% patchouli alcohol compound. The qualitative analysis by using thin layer chromatography (TLC) showed that there were similarities in the number of spot and Rf value for each spot of ex vitro and in vitro patchouli oil.

scholarly journals TDZ Plays Key Role in Shoot Regeneration from Different Explants of Picrorhiza kurroa: An Endangered Medicinal Herb of Western Himalayas

2019 ◽  
pp. 1-9 ◽  
Author(s):  
Vanita Patial ◽  
Amita Bhattacharya
Keyword(s):  
Natural Habitat ◽  
Basal Medium ◽  
In Vitro Rooting ◽  
Western Himalayas ◽  
Picrorhiza Kurroa ◽  
Regeneration Potential ◽  
Root Explants ◽  
Shoot Number ◽  
Endangered Medicinal Herb

Picrorhiza kurroa plants were collected from its natural habitat. In vitro plants were raised from the leaves of high yielding collection screened in an earlier study. Leaves, roots and internodal segments were cultured for 15 days. The effect of thidiazuron (1-phenyl-3-(1, 2, 3- thiadiazol-5-yl) urea; TDZ) pretreatment for 15 days on regeneration potential of different explants viz. leaves, roots and internodes of Picrorhiza kurroa was studied. Regeneration potential varied significantly with the type of explant. Regeneration response of 100% with 46.25 shoots per explant was obtained in leaf segments of 2.0 cm length pretreated with 0.5 µM TDZ for 15 days and then transferred to 2.32 µM kinetin (Kn) containing MS basal medium. In case of root explants maximum shoot number (17.12) was obtained on 0.5 µM TDZ pretreated for 15 days and then to 3.64 µM Kn. Maximum shoots per explants (12.33) were obtained in case of internodes pretreated with 0.5 µM TDZ for 15 days and transferred to 1.16 µM Kn. Regenerated shoots from different explants developed in vitro rooting on MS basal medium within 7-8 days. Conclusively, an efficient and repeatable protocol for rapid regeneration from different explants and in vitro rooting has been developed in P. kurroa which can be effectively used for its conservation.

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