The Effect of Heavy Metals on In Vitro Adventitious Shoot Production and Bacoside A Content in Bacopa Monnieri (L)

2017 ◽  
Vol 14 (4) ◽  
pp. 1-10
Author(s):  
Poornananda Madhava Naik Manoj Godbole
Keyword(s):  
Heavy Metals ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Dry Weight ◽  
Bacopa Monnieri ◽  
Optimum Number ◽  
Bacoside A ◽  
Shoot Production

In the present study the effect of heavy metals (MnSO 4, ZnSO 4 and CuSO 4 ) on adventitious shoot regeneration, biomass and bacoside A accumulation in Bacopa monnieri was carried out. The leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg l -1 kinetin (Kin) with varying concentration of heavy metals (Control: Mn-0.10 mM, Zn-0.03 mM and Cu- 0.0001mM; Mn: 0.20, 0.40, 0.80 and 1.60 mM; Zn: 0.06, 0.12, 0.24 and 0.48 mM; Cu: 0.02, 0.05, 0.10 and 0.20 mM). Optimum number of adventitious shoots (123.50 shoot/explants), fresh weight (3.826 g) and dry weight (0.226 g) of Bacopa monnieri were obtained in the medium with 0.12 mM Zn concentration. The highest production

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals Adventitious Shoot Regeneration and Rooting of Prunus serotina In Vitro Cultures

HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Ana Carolina Espinosa ◽  
Paula M. Pijut ◽  
Charles H. Michler
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Prunus Serotina ◽  
Naphthaleneacetic Acid ◽  
Eastern United States ◽  
Continuous Darkness ◽  
Number Of Shoots

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μm 6-benzylaminopurine (BA), 0.49 μm indole-3-butyric acid (IBA), and 0.29 μm gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μm thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μm naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μm BA in combination with 0, 0.54, 1.07, or 5.37 μm NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μm TDZ plus 0.54 μm NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μm TDZ plus 1.07 μm NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μm IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μm IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.

scholarly journals Adventitious Shoot Production from a Vireya Hybrid of Rhododendron

HortScience ◽  
1991 ◽  
Vol 26 (5) ◽  
pp. 594-596 ◽  
Author(s):  
Giovanni Iapichino ◽  
Tony H.H. Chen ◽  
Leslie H. Fuchigami
Keyword(s):  
Acetic Acid ◽  
Adventitious Shoots ◽  
Fold Increase ◽  
Adventitious Shoot ◽  
In Vitro Rooting ◽  
Axillary Shoots ◽  
Shoot Production ◽  
Shoot Tip Explants ◽  
Indole 3 Acetic Acid

An efficient adventitious shoot production protocol has been developed for Rhododendron laetum × aurigeranum. Shoot tips taken from greenhouse-grown plants were cultured on Anderson's medium supplemented with 74 μM 2iP. Axillary shoots were excised and cultured on medium containing 23 μM IAA and 74 μM 2iP. After 6 months, brown callus developed at the cut surfaces of the shoot-tip explants. This callus produced many adventitious shoots (up to 70 per explant). Clusters of adventitious shoots were divided, subculture, and continued to proliferate shoots. An estimated 1600-fold increase in the number of shoots could be readily achieved in 6 months. In vitro rooting of adventitious shoots was accomplished in 4 weeks. Seventy-three percent of shoots rooted on 1/4 strength Anderson's medium supplemented with 28 μm IAA. Plantlet survival was 100%3 weeks after transfer to soil. Chemical names used: 1-H-indole-3-acetic acid (MA); N-(3 -methy1-2-butenyl) -1H-purine-6 amine (2iP).

scholarly journals (291) Adventitious Shoot Production and Transformation of Euonymus alata

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1081C-1081
Author(s):  
Alan G. Smith ◽  
Elizabeth S. Zimmermann
Keyword(s):  
Butyric Acid ◽  
Adventitious Shoots ◽  
Adventitious Shoot ◽  
Kanamycin Resistance ◽  
Naphthaleneacetic Acid ◽  
Landscape Plant ◽  
Shoot Production ◽  
Four Levels ◽  
Number Of Shoots

Euonymus alata is an attractive landscape plant that has been reported to be an invasive species. Genetic modification through transformation is a method of reducing its invasiveness by producing sterile cultivars having limited or no seed production. A critical step in Agrobacterium-mediated gene transfer is the production of adventitious shoots. E. alata internodes and leaves from in vitro cultures were tested for adventitious shoot production on 16 plant growth regulator combinations: four levels of 6-benzylamino purine (BA) and three auxin treatments [0.5 or 0.25 mg·L-1 indole-3-butyric acid and 0.1 mg·L-1 naphthaleneacetic acid (NAA)], as well as no auxin. The optimal BA levels were found to be 0.5 or 1.0 mg·L-1 for maximizing the number of explants forming shoots and for producing the greatest number of shoots per explant. Culturing on NAA gave the greatest number of shoots per explant with both 0.5 and 1.0 mg·L-1 BA. Shoot production from internode segments was markedly superior to leaves. An initial dark treatment of 10 days did not influence shoot production. Using 1.0 mg BA with 0.1 mg·L-1 NAA, E. alata internodes were transformed with A. tumefaciens EHA105 carrying Kanamycin resistance and β-glucuronidase genes. Transformed shoots were selected on 30 mg·L-1 Kanamycin. Of the 36 shoots produced, 16 were confirmed to be transformed by β-glucuronidase histochemistry. Treatment with rooting powder containing indole-3-butyric acid did not aid rooting of shoots, but after 3 months in soil in high humidity, 21 of 24 E. alata shoots from tissue culture were rooted and acclimated.

scholarly journals Adventitious Shoot and Plantlet Formation from Cultured Apple Leaf Explants

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 629f-629
Author(s):  
Karim H. Al-Juboory ◽  
Jabar H. Al-Niami
Keyword(s):  
Tissue Culture ◽  
In Vitro Culture ◽  
Malus Domestica ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Adventitious Shoot ◽  
Plantlet Formation ◽  
Petri Dishes

Leaves of wild apple (Malus domestica Borkh) were excised from in vitro grown shoots transversely cut into halves and plated onto petri dishes containing regeneration media. Cultures were kept in the dark for three weeks before adventitious shoots were observed. Callus from leaf explants produced adventitious shoots after 3 months of in vitro culture. Callus were cultured on Nitsch and Nitsch medium supplemented with a range of BA (0.0–2.0 μm) and NAA (0.0–10 μm). BA at 10 μm combined with NAA (0.5 μm) proved most effective for stimulating shoot proliferation of cultured apple. Plantlets from tissue culture were easily transferred to the greenhouse environment.

Effects of sucrose and pH levels on in vitro shoot regeneration from leaf explants of Bacopa monnieri and accumulation of bacoside A in regenerated shoots

2009 ◽  
Vol 100 (2) ◽  
pp. 235-239 ◽  
Author(s):  
Poornananda Madhava Naik ◽  
Shirugumbi Hanamanthagouda Manohar ◽  
Nagella Praveen ◽  
Hosakatte Niranjana Murthy
Keyword(s):  
Shoot Regeneration ◽  
Leaf Explants ◽  
Bacopa Monnieri ◽  
Bacoside A ◽  
In Vitro Shoot Regeneration

scholarly journals Adventitious Shoot Formation on Hypocotyl Explants of Antirrhinum majus L.

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 477B-477
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Adventitious Shoots ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Antirrhinum Majus ◽  
Fluorescent Light ◽  
Adventitious Shoot Formation ◽  
Hypocotyl Explants ◽  
Shoot Production ◽  
Dark Regime

One-centimeter hypocotyl explants from 2-week-old Antirrhinum majus L. (snapdragon) seedlings germinated and grown in vitro under 12-h cool-white fluorescent light and 12 h dark or 24 h dark were placed on Murashige and Skoog (MS) medium containing 0, 0.44, 2.22, 4.44, 8.88, or 44.4 μM N6-benzyladenine (BA). Cultures were maintained under the light/dark regime at 25°C. After 2 weeks, adventitious shoots were counted. A shoot was considered adventitious and counted if a stem and leaf developed. Shoots developed along the entire length of the hypocotyl sections. Mean shoot production per hypocotyl explant ranged from 2.4 to 6.1 shoots when seedlings were germinated and grown in 24 h darkness and 2.2 to 10.9 shoots when started in the light/dark regime. Highest shoot counts were attained /from hypocotyl explants when seedlings were germinated and grown under the light/dark regime for 2 weeks and transferred to 2.22, 4.44, or 8.88 μM BA. Shoot development appeared normal at the 2.22 and 4.44 μM level, while at 8.88 μM BA, development was slightly abnormal along with slightly more callus production.

scholarly journals An Efficient Tissue Culture Regeneration System for Georgia Plume, Elliottia racemosa, a Threatened Georgia Endemic

HortScience ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 447-453 ◽  
Author(s):  
Seong Min Woo ◽  
Hazel Y. Wetzstein
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Dry Weight ◽  
Medium Composition ◽  
Induction Medium ◽  
Ex Vitro ◽  
Regeneration System

Georgia plume, Elliottia racemosa Muhlenb. ex. Elliott, is an extremely rare small tree or shrub endemic to Georgia, which is being severely affected by habitat loss and lack of sexual recruitment. In vitro plant regeneration of Georgia plume has not been previously reported and may be a method for the conservation and propagation of this threatened species. Studies evaluated the effects of sterilization methods, explant types, medium composition, and light environment on plant regeneration. An efficient plant regeneration system was developed in which adventitious shoot buds were induced using young, expanding leaf explants placed on an induction medium supplemented with 10 μm thidiazuron and 5 μm indole-3-acetic acid with Gamborg's B5 salts. Shoot elongation was promoted on a medium with 25 μm (2-isopentenyl) adenine incorporated into Woody Plant Medium. In vitro rooting studies evaluated continuous and pulse auxin treatments and ex vitro rooting trials after KIBA (indole-3-butric acid, potassium salt) dips. A 5-day pulse treatment on 100 or 150 μm indole-3-butyric acid produced ≈90% rooting of shoots with greater shoot and root dry weight than other pulse times. High rooting frequencies were obtained under in vitro and ex vitro conditions with over 85% survival of plantlets transferred to greenhouse conditions. The culture protocol was found to be effective with material collected from mature specimens in the wild from divergent populations. Tissue culture appears to be a promising approach for the propagation and conservation of this rare and threatened plant.

scholarly journals Regenerating Plants from in Vitro Culture of Black Locust Cotyledon and Leaf Explants

HortScience ◽  
1993 ◽  
Vol 28 (9) ◽  
pp. 942-945 ◽  
Author(s):  
I. Arrillaga ◽  
S.A. Merkle
Keyword(s):  
Shoot Regeneration ◽  
Black Locust ◽  
Robinia Pseudoacacia ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Regeneration Frequency ◽  
Adventitious Shoot Formation

A protocol to achieve efficient plant regeneration from juvenile black locust (Robinia pseudoacacia L.) explants is described. Direct adventitious shoots were induced from cotyledon explants on woody plant medium containing 22.2 μm BA and 0.4 μm 2,4-D. Shoots developed and new shoots were induced when the explants were transferred to medium without growth regulators. The effect of dark incubation on shoot regeneration from cotyledons indicated that 15 days of darkness resulted in a high regeneration frequency (91.7%). Adventitious shoot formation also was induced from sections of in vitro-derived leaves cultured in darkness on Murashige and Skoog medium supplemented with 4.4 μm BA and 24.6 μm IBA. A shoot regeneration frequency of 89% was obtained when explants were subcultured on a medium containing 4.4 μm BA and 0.5 μm IAA. Shoots were rooted on Schenk and Hildebrandt medium with or without IBA. Plantlets were acclimatized and grown in the greenhouse. Chemical names used: N -(phenylmethyl)-1H -purin-6-amine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA).

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaves of Formosan Sweetgum (Liquidambar formosana L.)

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 721-723 ◽  
Author(s):  
L. Xu ◽  
G.F. Liu ◽  
M.Z. Bao
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
High Regeneration

Plantlets were regenerated from in vitro-grown leaf explants of five genotypes of Liquidambar formosana on WPM basal medium supplemented with different concentrations of TDZ and NAA. With the addition of 0.27 μm NAA, regeneration efficiency was increased by 2- to 4-fold over that with TDZ alone. Lower concentrations of TDZ (0.45–2.27 μm) were beneficial for regenerating shoot clusters. Four genotypes (P2, P6, P9, and P11) showed high regeneration rates (up to 90%), whereas genotype P13 showed a low capability for shoot regeneration on all media tested (<35%). For all five genotypes, the optimum medium for inducing adventitious shoots was WPM supplemented with 1.14 μm TDZ and 0.27 μm NAA, on which regeneration rate ranged from 72.6% to 89.5% and adventitious shoot clusters per regenerating leaf explant ranged from 2.63 to 3.11 in four genotypes (P2, P6, P9, and P11), while for P13, the regeneration rate and number of shoot clusters per regenerating explant were 23% and 1.39, respectively. Transfer of shoot clusters to WPM basal medium containing 0.54 μm NAA, 2.22 μm BA, and 1.44 μm GA3, resulted in shoot elongation. All the elongated shoots were rooted on WPM supplemented with 9.84 μm IBA, and plantlets were transplanted to soil successfully. Chemical names used: 6-benzyladenine (BA), gibberellic acid (GA3), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), plant growth regulator (PGR), thidiazuron (TDZ), woody plant medium (WPM).

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