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 In vitro Multiple Shoot Regeneration from Petunia hybrida

2019 ◽  
Vol 7 (10) ◽  
pp. 1554
Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir
Keyword(s):  
Petunia Hybrida ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Length ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.

scholarly journals In Vitro Long-Term Cultures of Papaya (Carica Papaya L.).

2021 ◽  
Author(s):  
Jose Javier Regalado González ◽  
Manuel López Granero ◽  
Carlos Lopez Encina
Keyword(s):  
Morphological Characteristics ◽  
Basal Medium ◽  
Multiplication Rate ◽  
Ms Medium ◽  
High Quality ◽  
Poor Growth ◽  
Average Multiplication ◽  
Half Strength

Abstract We present the data on proliferation corresponding to 10 years of continuous incubation in vitro of papaya shoots, and propose a reliable method for long-term micropropagation for papaya, using two types of explants: Microshoots from somatic embryos, and from axillary buds of papaya. Three different media were assayed. The proliferation medium (PPRM) allowed to maintain papaya shoots under continuous proliferation during 20 years, maintaining a consistent behaviour. Most of the shoots developed in PPRM rooted during the incubation, and after acclimated easily, maintaining the morphological characteristics of the parental plants, flowering and setting fruits normally. The PPRM medium consist in MS medium supplemented with NAA (0.1 mg l-1), BA (0.5 mg l-1), GA3 (0.5 mg l-1) and Adenine sulphate (40 mg l-1). The average multiplication rate was higher than 20 shoots per explant along the long-term assay. The elongation medium (PELM), was designed to recover shoots with a poor growth, and allowed the development of high quality shoots ready for rooting, and consist in a MS basal medium supplemented with NAA (0.1 mg l-1), Kin (0.5 mg l-1) and GA3 (1 mg l-1). The rooting medium (PROM) was designed to induce high quality roots from non-rooted shoots and consist in a half strength MS medium plus IBA (1mg l-1). On PROM, agar can be exchanged for expanded vermiculite. Acclimation took place inside an acclimatization tunnel under progressive hydric stress. After 4 weeks, the plant recovery rate was 90% for plants maintained under continuous proliferation during ten years.

scholarly journals Effect of Seed Orientation and Medium Strength on In vitro Germination of Pterocarpus marsupium Roxb.

2013 ◽  
Vol 5 (4) ◽  
pp. 476-479 ◽  
Author(s):  
Yogeshwar MISHRA ◽  
Rimi RAWAT ◽  
Brajesh NEMA ◽  
Fatima SHIRIN
Keyword(s):  
Shoot Length ◽  
Ms Medium ◽  
Root Number ◽  
Pterocarpus Marsupium ◽  
Shoot Number ◽  
Medium Strength ◽  
Half Strength ◽  
Varied Response ◽  
Strength Medium

A method was developed for optimization of In vitro germination of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. The seeds inoculated in different orientation on different strengths of MS medium without any hormones showed varied response to the seed positions. The seeds inoculated on half strength medium in horizontal position recorded to produce a maximum germination (78.23%), shoot number (0.86) and root number (7.99). However, a maximum of shoot length of 3.67 cm was recorded in the quarter strength medium in the seeds inoculated in vertical down position, which was significantly higher than other media strength and positions. Our results indicate that the seed orientation including medium strength have tremendous effect on germination and seeds inoculated horizontally on half strength MS medium can be utilized to enhance In vitro seed germination of Pterocarpus marsupium.

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).

Seed germination, micropropagation from adult and juvenile origin explants and address of hyperhydricity of the Cretan endemic herb Calamintha cretica

2020 ◽  
Vol 48 (3) ◽  
pp. 1504-1518
Author(s):  
Georgia VLACHOU ◽  
Maria Maria PAPAFOTIOU ◽  
Konstantinos Bertsouklis
Keyword(s):  
Shoot Proliferation ◽  
Shoot Length ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Optimum Range ◽  
Shoot Number ◽  
Half Strength ◽  
One Year ◽  
Cardinal Temperatures

The optimum range of temperature for germination (96-100%) of Calamintha cretica, an herb with potential pharmaceutical and horticultural uses, was 15 to 20 °C, with 10 and 30 °C cardinal temperatures. Storage up to one year did not affect germination. The effect of zeatin (ZEA), 6-benzyladenine (BA), kinetin, and 6-γ-γ-(dimethylallylamino)-purine added in MS medium at concentrations from 0.0 to 8.0 mg L-1 was tested for shoot proliferation of both adult- and seedling-origin nodal explants at first- and sub-culture. Both explant types responded similarly during in vitro culture. At cytokinin concentrations up to 1 mg L-1 explant response was high (over 85%) but shoot number per explant was low (1.2-2.2). Increasing cytokinin from 2.0 to 8.0 mg L-1 resulted to an analogous decrease of explant response and shoot length, and an increase of shoot number, particularly when ZEA or BA was used (5.0-6.6 shoots per explant, 0.5-1.0 cm long) with simultaneous though increase of hyperhydricity (up to 50%). The addition of 0.1 mg L-1 naphthaleneacetic acid into the 8.0 mg L-1 BA medium almost eliminated hyperhydricity and increased explant response, while the increase of agar concentration from 8.0 to 12.0 g L-1 eliminated hyperhidricity and induced the highest shoot proliferation (93-95% explant response, 11.2-12.3 shoots per explant, 0.8-1.0 cm long). Microshoots and microshoot clusters rooted (88-96%) on half-strength MS medium either hormone free or supplemented with 1 to 4 mg L-1 indole-3-butyric acid. Plantlets survived at 80% to 100% after ex vitro acclimatization in peat: perlite 1:1 (v/v).

scholarly journals Mass propagation of Bambusa bambos (L.) Voss through in vitro culture

2017 ◽  
Vol 5 (2) ◽  
pp. 15-26 ◽  
Author(s):  
Raihan I Raju ◽  
Shyamal K Roy
Keyword(s):  
In Vitro Culture ◽  
Basal Medium ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Surface Sterilization ◽  
Half Strength

Protocol for mass propagation of Bambusa bamboos (L.) Voss was developed through in vitro culture. Nodal segments containing pre-existing axillary bud, after surface sterilization, were inoculated on liquid Murashige and Skoog’s (MS) basal medium containing different concentrations and combinations of cytokinins (BAP, TDZ and Kn). The highest direct shoot induction (90%) was obtained in the MS liquid medium supplemented with 2.0 mg/l BAP and 1.0 mg/l TDZ with maximum average number of shoots (3.14 ± 0.06) per explant. Highest shoot multiplication (16.58 ± 0.24 shoots per culture) with highest average shoot length (9.21 ± 0.13 cm) was obtained when in vitro raised shoots were cultured in gelrite gelled MS medium in conjunction with 2.0 mg/l BAP and 1.0 mg/l TDZ. Incorporation of 10% coconut water with 4% sucrose in the above mentioned medium resulted satisfactory shoot growth and development with an average 26.7 ± 0.60 shoots per culture. For root induction, in vitro raised shoots were divided into clumps of 4-5 shoots in each clump and transferred onto both liquid and gelled half-strength MS medium containing different concentrations and combinations of auxins (IBA and NAA). Maximum rooting (86.67%) was achieved in half-strength of MS medium fortified with 2.5 mg/l IBA and 2.5 mg/l NAA with an average 8.72 ± 0.42 root per shoot. The rooted plantlets were then transferred to polybags containing garden soil, sand and compost mixture with 1:1:1 ratio. After a month the hardened plantlets were then transferred to the larger pots containing garden soil and compost with 1:1 ratio for sufficient growth and finally transplanted to the field. In this process, the highest 100% survivability was recorded from well-established rooted plantlets. The regenerated plants showed well developed root and shoot systems in field condition.Jahangirnagar University J. Biol. Sci. 5(2): 15-26, 2016 (December)

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 A Protocol for Rapid Clonal Propagation and Microrhizome Production of Curcuma caesia Roxb. (Zingiberaceae): A Critically Endangerd Medicinal Plant of North East India

2020 ◽  
Author(s):  
Indrani Sarma ◽  
A. C. Deka ◽  
T. C. Sarma
Keyword(s):  
Medicinal Plant ◽  
Basal Medium ◽  
Shoot Multiplication ◽  
Cost Effective ◽  
Northeast India ◽  
Ms Medium ◽  
North East India ◽  
North East ◽  
Best Response

Background: Curcuma caesia Roxb. (Zingiberaceae) is a rare, critically endangered medicinal plant of Northeast India. The rhizome of the plant is famous for its significant medicinal properties. Methods: Various concentrations of plant growth regulators in Murashige and Skoog (MS) medium were tried in the study using rhizome bud as explant for development of an efficient cost effective protocol for in vitro mass multiplication and microrhizome induction of Curcuma caesia. Result: Shoot multiplication and plant generation was achieved from freshly sprouted shoots of Curcuma caesia on Murashige and Skoog’s medium supplemented with different phytohormones. The best response for shoot multiplication (23.80±0.51shoots/explant) was obtained within 8 weeks in MS medium supplemented with BAP (1.0 mgl-1) and 2,4-D (0.25 mgl-1). Microrhizomes were induced at the base of the in vitro derived shoots upon transferred to medium containing various combinations and concentrations of sucrose and BAP and grown under varying photoperiod. Half strength of MS basal medium containing BAP (1.0 mg/l) and 9% sucrose was found to be optimum for induction of large sized microrhizome within 45 days of incubation under 16 hrs of photoperiod.

scholarly journals Development of in vitro propagation system for Atriplex halimus L.

2014 ◽  
Vol 20 (3-4) ◽  
Author(s):  
O. Aldahhak ◽  
S. Zaid ◽  
A. M. Abdul-Kader
Keyword(s):  
In Vitro Selection ◽  
Basal Medium ◽  
Optimal Growth ◽  
Multiplication Rate ◽  
Ms Medium ◽  
In Vitro Shoots ◽  
Shoot Number ◽  
Nacl Concentration ◽  
Selection For

Explants excised from adult shrubs were surface sterilized and cultured on Murashige and Skoog (MS) basal medium in the presence of plant growth regulators (PGRs) at different concentrations. A high multiplication rate of 7.2-fold was achieved every four weeks on MS medium supplemented with 4.44 μM BA, 0.49 μM IBA and 0.58 μM GA3. Rooting was achieved with 73% efficiency within 2-4 weeks on agar-gelled MS basal medium free of PGRs. Rooted plantlets were gradually acclimatized to field conditions over 5-6 weeks with 65% efficiency. For in vitro selection for salt tolerance, MS medium was supplemented with increasing concentrations of NaCl ranging between 25 and 1000 mM. This study has demonstrated that in vitro shoots could tolerate up to 600 mM NaCl with optimal growth at 200 mM, while higher concentrations of NaCl affected growth negatively. Growth and shoot number decreased with increasing NaCl concentration with all plantlets died at 1000 mM NaCl.

Evaluation of explants for in vitro propagation of Citrus indica Tanaka - An Endangered Species

2020 ◽  
Vol 30 (1) ◽  
pp. 87-96
Author(s):  
Shabaree YA Sangma ◽  
Lolly S Pereira ◽  
JC Dang ◽  
Binu Mathew
Keyword(s):  
Leaf Disc ◽  
Shoot Length ◽  
Shoot Tip ◽  
Root Tip ◽  
Nodal Segment ◽  
Ms Medium ◽  
Shoot Bud ◽  
Shoot Number ◽  
Half Strength

Of the five explants viz., shoot tip, nodal segment, leaf disc, cotyledon and root tip excised from exegenic seedlings of Citrus indica Tanaka shoot tip cultured in MS supplemented with 0.5 mg/l of BAP generated shoots within 4.74 days, exhibiting highest percentage of response (85.82%) with highest number of shoots (8.9) and shoot length (3.04 cm). On the other hand nodal segment cultured in MS supplemented with 1.0 mg/l BAP showed 80% response in 5.16 days with a shoot number of 5.41 and shoot length of 2.43 cm. Cotyledon explants inoculated on MS supplemented with1.0 mg/l of TDZ produced shoots in 20 days with the highest response of 69.88%, with 3.77 shoots per cotyledon and shoot length of 2.03cm. Viable callus was obtained from leaf disc cultured on half strength MS medium with less Ca++ with 2, 4-D 0.5 mg/l + Kn 0.25 mg/l. This callus when inoculated on half strength MS medium with Kn 1.5 mg/l showed highest shoot bud proliferation of 66.66% with 10.06 shoots per callus. Root tip explant failed to produce any shoots. In vitro raised shoots of Citrus indica when cultured on half strength MS medium supplemented with NAA (1.0 mg/l) showed 80 % rooting in 5.66 days, with highest number of roots (6.16 per shoot) and longest root (3.78cm). Ninety per cent of in vitro rooted plantlets of Citrus indica survived in open conditions. Plant Tissue Cult. & Biotech. 30(1): 87-96, 2020 (June)

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