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

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

Plants ◽

10.3390/plants9060755 ◽

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.

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Plant Regeneration through Protocorm-like Bodies Induced from Nodal Explants of Syngonium podophyllum ‘White Butterfly’

HortScience ◽

10.21273/hortsci.43.7.2129 ◽

2008 ◽

Vol 43 (7) ◽

pp. 2129-2133 ◽

Cited By ~ 5

Author(s):

Jin Cui ◽

Juanxu Liu ◽

Min Deng ◽

Jianjun Chen ◽

Richard J. Henny

Keyword(s):

Adventitious Shoots ◽

Leaf Explants ◽

Basal Medium ◽

Shoot Culture ◽

Nodal Explants ◽

Induction Medium ◽

Naphthalene Acetic Acid ◽

Ex Vitro ◽

Petiole Explants

Syngonium podophyllum ‘White Butterfly’, one of the most popular ornamental foliage plants, is propagated almost exclusively through in vitro shoot culture. Ex vitro rooting, however, has been associated with severe Myrothecium leaf spot (Myrothecium roridum Tode ex Fr.). The objective of this study was to establish a method for regenerating well-rooted plantlets before ex vitro transplanting. Leaf and petiole explants were cultured on a Murashige and Skoog (MS) basal medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ), 6-benzyladenine (BA), or N-isopentenylaminopurine (2iP) with α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. Calli formed from leaf explants cultured on the basal medium supplemented with CPPU or TDZ with 2,4-D or with NAA as well as from petiole explants cultured on the medium supplemented with BA, CPPU, or TDZ with 2,4-D or NAA. The calli, however, failed to differentiate, and shoot organogenesis did not occur. Culture of nodal explants on the MS basal medium supplemented with 9.84 μm 2iP, 8.88 μm BA, 8.07 μm CPPU, or 9.08 μm TDZ with 2.26 μm 2,4-D resulted in the formation of protocorm-like bodies, adventitious shoots, and subsequently well-rooted plantlets. MS basal medium supplemented with 19.68 μm 2iP and 1.07 μm NAA resulted in the highest percentage (92.9%) of nodal explants producing protocorm-like bodies and an average of 16.9 well-rooted plantlets per nodal explant. Adventitious shoots were able to root in the initial induction medium, but better root development occurred after shoots with protocorm-like bodies were transferred onto MS basal medium supplemented with 9.84 μm 2iP and 2.69 μm NAA. Regenerated plantlets were stable and grew vigorously with 100% survival rates after ex vitro transplanting to a container substrate in a shaded greenhouse.

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Adventitious Shoot Regeneration and Rooting of Prunus serotina In Vitro Cultures

HortScience ◽

10.21273/hortsci.41.1.193 ◽

2006 ◽

Vol 41 (1) ◽

pp. 193-201 ◽

Cited By ~ 37

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.

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Plant Regeneration from Leaf of Streptocarpus × hybridus Voss. Cultured In Vitro

HortScience ◽

10.21273/hortsci.39.4.813a ◽

2004 ◽

Vol 39 (4) ◽

pp. 813A-813

Author(s):

Maria Cantor* ◽

Rodica Pop ◽

Ioana Pop

Keyword(s):

Shoot Regeneration ◽

Total Duration ◽

Leaf Explants ◽

Basal Medium ◽

Adventitious Shoot ◽

Direct Shoot Regeneration ◽

Limited Scale ◽

The Media ◽

Tissue Culture Propagation

The Streptocarpus is propagated ease vegetatively from leaf cuttings all year round, but is grown on a very limited scale commercially in Romania. Successful protocol for direct shoot regeneration from in vitro Cape primrose (Streptocarpus × hybridus Voss.) leaf explants has been developed. The ease of tissue culture propagation can promote Streptocarpus production and facilitated the rapid introduction of this new species. Adventitious shoot regeneration was inducted in vitro on MS basal medium, using different concentration of NAA (1, 1.5, 2 mg·L-1) and cyokinin TDZ (0.1, 0.5, 1 mg·L-1). High frequency regeneration was obtained from leaves when cultured in the media supplemented with 1 mg·L-1 NAA plus 0.5 mg·L-1 TDZ and the percent of regeneration resulted is between 70% to 100%. Complete plantlets were acclimatized and successfully transplanted to glasshouse conditions. The total duration of the cycle from leaf explants through complete plantlets was 10 weeks.

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Sodium Nitroprusside Stimulates Micropropagation and TDZ Induces Adventitious Shoots Regeneration in Red Flesh Apple Malus niedzwetzkyana Koehne Dieck ex

Journal of Horticultural Research ◽

10.2478/johr-2019-0014 ◽

2019 ◽

Vol 27 (2) ◽

pp. 23-30

Author(s):

Nooshin Kazemi ◽

Maryam Jafarkhani Kermani ◽

Ali Akbar Habashi

Keyword(s):

Sodium Nitroprusside ◽

Shoot Regeneration ◽

Exposure Time ◽

Adventitious Shoots ◽

Leaf Explants ◽

Adventitious Shoot ◽

Interactive Effects ◽

Naphthalene Acetic Acid ◽

Adventitious Shoot Regeneration ◽

Basal Section

AbstractThe aim of the present investigation was to optimize protocols for micropropagation and adventitious shoot regeneration from leaf explants of two wild ecotypes of red flesh apple Malus niedzwetzkyana for future breeding programs. At the proliferation stage, different concentrations of sodium nitroprusside (SNP) and triacontanol (TRIA) were compared. To optimize shoot regeneration from leaf explants, interactive effects of 1-phenyl-3-(1,2,3-thidiazol-5-yl)-urea – thidiazuron (TDZ), indole-3-butyric acid (IBA) and two explant types were investigated. At rooting stage, the effect of exposure time of microshoots to darkness and exposure time to different concentrations of IBA and α-naphthalene acetic acid (NAA) were compared. The results showed that SNP affected the growth rate significantly and the maximum multiplication rates per explant (9.6 in the first ecotype and 8.8 in the second) were produced in the Quoirin and Lepoivre medium containing 17 SNP µM, in addition to 4 µm 6-benzylaminopurine (BAP) and 3 µm gibberellic acid (GA3). IBA and TDZ affected the adventitious shoot regeneration from leaf explants significantly, the highest number of regenerated shoots (18.3 per explant) was obtained from the basal section of the leaves cultured on the medium containing 2 μM IBA and 15 μM TDZ. At rooting stage, the maximum rooting (88.6%) was obtained in the result of one weak exposure to darkness on medium containing 3 μM IBA.

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Regeneration of Chlorophytum amaniense ‘Fire Flash’ Through Indirect Shoot Organogenesis

HortScience ◽

10.21273/hortsci.46.3.466 ◽

2011 ◽

Vol 46 (3) ◽

pp. 466-469

Author(s):

Jin Cui ◽

Juanxu Liu ◽

Jianjun Chen ◽

Richard J. Henny

Keyword(s):

Callus Formation ◽

Adventitious Shoots ◽

Leaf Explants ◽

Basal Medium ◽

Culture Method ◽

Shoot Formation ◽

Photon Flux ◽

Naphthalene Acetic Acid ◽

Foliage Plant

Chlorophytum amaniense Engl. ‘Fire Flash’ is a popular exotic ornamental foliage plant as a result of its unique coral-colored midribs and petioles and tolerance to interior low light levels. Currently, demand for propagative materials exceeds the availability of seeds. This study was intended to develop an in vitro culture method for rapid propagation of this cultivar. Leaf and sprouted seed explants were cultured on a Murashige and Skoog basal medium supplemented with different cytokinins with 1.1 μM α-naphthalene acetic acid (NAA) or 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf explants showed poor responses in callus production and no adventitious shoots were obtained. Callus formation frequencies from sprouted seeds were 71% and 85% when induced by 9.8 μM N6-(2-isopentyl) adenine (2iP) with 1.1 μM NAA and 9.1 μM N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with 1.1 μM NAA, respectively. Adventitious shoots occurred after the induced calluses were subcultured on the same concentrations of TDZ or 2iP with NAA. Shoot formation frequencies from calluses cultured on TDZ with NAA and 2iP with NAA were 92% and 85%, and the corresponding mean shoot numbers were 37 and 31 per piece of callus (1 cm3), respectively. Adventitious shoots rooted at 100% after transferring to the basal medium containing 4.4 μM 6-benzylaminopurine (BA) with 2.7 μM NAA. Plantlets, after transplanting to a soilless substrate were easily acclimatized in a shaded greenhouse under a photosynthetic photon flux (PPF) density of 200 μmol·m−2·s−1. Regenerated plants grew vigorously without undesirable basal branching or distorted leaves. This newly established regeneration method can provide the foliage plant industry with a means for rapidly propagating ‘Fire Flash’ liners in a year-round fashion.

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Comparative Organogenic Response of Six Clonal Apple Rootstock Cultivars

HortScience ◽

10.21273/hortsci.51.3.272 ◽

2016 ◽

Vol 51 (3) ◽

pp. 272-278 ◽

Cited By ~ 1

Author(s):

Qingrong Sun ◽

Meijuan Sun ◽

Hongyan Sun ◽

Richard L. Bell ◽

Linguang Li ◽

...

Keyword(s):

Shoot Regeneration ◽

Shoot Organogenesis ◽

Leaf Explants ◽

Basal Medium ◽

Medium Composition ◽

Apple Rootstock ◽

Malus Prunifolia ◽

Root Organogenesis ◽

High Regeneration

The organogenesis potential is different among cultivars and must be optimized for individual genotype. Shoot organogenesis capacity from in vitro leaves and root organogenesis capacity of in vitro shoots in six clonal apple rootstock cultivars were compared. The shoot organogenesis capacity was highly genotype dependent. ‘GM256’ was found to be the most responsive genotype for shoot regeneration from leaf explants among the cultivars, showing high regeneration percentage on all tested media. The effects of basal medium composition and cytokinins on shoot regeneration were different depending on rootstock genotype. Optimum regeneration occurred on Murashige and Skoog (MS) basal medium for ‘71-3-150’, and optimum regeneration occurred on Quoirin and Lepoivre (QL) basal medium for ‘60-160’ and ‘ПБ’. Thidiazuron (TDZ) was more effective than 6-benzylaminopurine (BA) for Malus prunifolia (Y), whereas TDZ and BA were not significantly different for the other cultivars. All rootstock cultivars showed high root organogenic capacity. The percentage of rooting reached more than 90% and the mean root number per plantlet ranged from three to five. The optimum rooting medium was different for different rootstock cultivars. Optimum root organogenesis occurred on half-strength QL medium for ‘GM256’ and ‘Y’, and for ‘ПБ’ and ‘JM7’ on one-quarter-strength MS medium.

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In vitro callus and shoot regeneration in Enterolobium cyclocarpum (Jacq.) Grised. - a fast timber yielding species

Notulae Scientia Biologicae ◽

10.15835/nsb12110604 ◽

2020 ◽

Vol 12 (1) ◽

pp. 74-89

Author(s):

Michael S. AKINROPO ◽

Benjamin E. AYISIRE ◽

Ejeoghene R. OGBIMI

Keyword(s):

Shoot Regeneration ◽

Leaf Explants ◽

Basal Medium ◽

Multiple Shoots ◽

Nodal Explants ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Cotyledon Explants ◽

Enterolobium Cyclocarpum

This study was conducted to investigate the in vitro callus induction and rapid shoot regeneration potential in Enterolobium cyclocarpum, a plant native to central Mexico but widely introduced into Africa. The leaf, stem and nodal explants of E. cyclocarpum were cultured on full strength Murashige and Skoog (MS) medium supplemented with different concentrations of Cytokinins - Benzyladenine (BA) and/or Kinetin and Auxins - Naphthalene acetic acid (NAA) and/or 2,4-Dichlorophenoxylacetic acid (2,4-D) each alone and in combination. The leaf explants did not respond to these treatments. The Nodal explants were best for caulogenesis, while the explant responses were in the order- nodal > stem > cotyledon for callogenesis in MS medium supplemented with BA and/or Kin combined with NAA and/or 2,4-D. The varied combinations induced white compact callus. The highest callus production was observed on MS medium supplemented with 2.7 µM NAA + 2.2 µM BA and 5.4 µM NAA alone. Nodal and cotyledon explants developed callus and multiple shoots on MS supplemented with a combination of cytokinin (BA and/or Kin.) and auxin (NAA and/or 2,4-D). The maximum number of 3.98 ± 0.37 and 2.1±0.11 shoots/explants were recorded for nodal and cotyledon explants on MS medium supplemented with a combination of 8.8 µM BA+2.7 µM NAA and 2.2µM BA+2.7 µM NAA respectively. On the basal medium, 10% of the excised shoots rooted successfully. Thus, this in vitro method can be exploited for conservation and mass propagation of this fast timber yielding tree and also utilized for embryogenesis studies.

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Factors Affecting Plant Regeneration from Leaf Tissues of Buddleia Species

HortScience ◽

10.21273/hortsci.42.7.1670 ◽

2007 ◽

Vol 42 (7) ◽

pp. 1670-1673 ◽

Cited By ~ 9

Author(s):

Wenhao Dai ◽

Cielo Castillo

Keyword(s):

Shoot Regeneration ◽

Basal Medium ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Dark Treatment ◽

Regeneration Rate ◽

Factors Affecting ◽

In Vitro Shoots ◽

Leaf Tissues

The effects of genotype, basal medium, plant growth regulator (PGR), dark treatment, and antibiotics on shoot regeneration of two Buddleia cultivars, B. davidii ‘Potters Purple’ and Buddleia ‘Lochinch’, were investigated. In vitro shoots were regenerated from leaf tissues in either Murashige and Skoog (MS) or woody plant medium (WPM) media supplemented with benzyladenine (BA). In general, more shoots were regenerated in WPM medium than in MS medium. Dark treatment for 3 to 5 weeks dramatically increased shoot regeneration. Addition of indole-3-butyric acid (IBA) or naphthalene acetic acid (NAA) significantly enhanced the regeneration rate and shoots of each explant. The maximum regeneration rate (100%) of B. davidii ‘Potters Purple’ was achieved when cultured in WPM containing 5 μm BA plus 5 μm IBA. The maximum regeneration rate (98.4%) of Buddleia ‘Lochinch’ was found in WPM supplemented with 20 μm BA plus 4 μm IBA. Carbenicillin at 250 to 500 mg·L−1 and cefotaxime at 125 to 250 mg·L−1, individually or combined, promoted shoot regeneration. Interactions between genotype and medium or PGRs were found. In vitro shoots were easily rooted in half-strength MS medium with or without NAA. Rooted plants were transferred to potting mix and grown in the greenhouse. This research will facilitate genetic improvement and fast propagation of Buddleia species using biotechnology.

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

HortScience ◽

10.21273/hortsci.28.9.942 ◽

1993 ◽

Vol 28 (9) ◽

pp. 942-945 ◽

Cited By ~ 6

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).

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