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

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.

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 721-723 ◽  
Author(s):  
L. Xu ◽  
G.F. Liu ◽  
M.Z. Bao

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).


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1077B-1077
Author(s):  
Wenhao Dai ◽  
Cielo Castillo ◽  
Victoria Magnusson

In vitro shoot cultures for two birch species, Asian white birch (Betula platyphylla) and paper birch (Betula papyrifera), were initiated from shoot tips of mature trees and maintained in MS (Murashige and Skoog) medium containing 3% sucrose and 5–10 μM (micromolar) benzyladenine (BA). The effect of such factors as genotype, basal medium, and plant growth regulator (PGR) on proliferation was investigated. Shoots were proliferated in both MS and woody plant medium (WPM) supplemented with different concentrations of thidiazuron (TDZ), BA, and kinetin (Kin). Two birch species responded differently to these factors. In general, more shoots were proliferated in WPM than in MS medium. The maximum proliferation rate of Asian white birch was achieved by being cultured in WPM containing 4–8 μM TDZ, while paper birch gave rise to the maximum proliferation rate in WPM supplemented with 20 μM BA. Interactions between genotype and medium or cytokinin were found. Shoots produced on media with TDZ had thick stems and small, dark green leaves. Microshoots can be rooted both in vitro and ex vitro with or without IBA treatment. Plants were regenerated from leaf tissues of Asian white birch. Adventitious shoots regenerated when in vitro leaves were cultured on WPM supplemented with 10–20 μM BA with 2-week dark treatment. The effect of genotype, PGR, and culture condition on in vitro regeneration of birch species is being tested.


1970 ◽  
Vol 18 (2) ◽  
pp. 173-179 ◽  
Author(s):  
T. Mallikadevi ◽  
P. Senthilkumar ◽  
S. Paulsamy

The in vitro regeneration of Plubago zeylanica exhibited that the callus was initiated in the basal medium containing BAP, NAA, 2, 4-D, and IBA.  The high amount (90%) of organic calli was induced in the basal medium supplemented with 2, 4-D, alone at 2.0 mg/l. In the subculture the adventitious shoot formation was prominently higher (83%) in the basal medium containing BAP, and NAA at 3.5 and 0.3 mg/l, respectively. IAA (1.0 mg/l)effectively produced higher percen-tage (90) of roots and root growth. After sequential hardening, survivability rate was observed to be significantly higher (80%) in the hardening medium containing garden soil, sand and vermicompost in the ratio of 1 : 1 : 1 by volume under greenhouse condition.  Key words: Plumbago zeylanica, In vitro regeneration, Medicinal plant D.O.I. 10.3329/ptcb.v18i2.3648 Plant Tissue Cult. & Biotech. 18(2): 173-179, 2008 (December)


HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 471B-471
Author(s):  
Agustin Huerta ◽  
Ramon Dolcet-Sanjuan

Adventitious shoots and viable plants were regenerated from bell pepper (Capsicum annuum L.) cultivars and dihaploid lines (DHLs) obtained from F1 hybrids via androgenesis (Dolcet-Sanjuan et al., in press). Hypocotil and cotyledon sections from in vitro-germinated seeds were used as explants. A modified MS medium (Murashige and Skoog, 1962) supplemented with IAA (0 to 3.2 μM) and BAP (0 to 100 μM) was used in a 3-week-long shoot primordia induction phase. Shoot elongation was best performed in the same basal medium, but supplemented with silver thiosulfate and GA3. Shoots were regenerated from eight selected DHLs (`C213', `C215', `C218', `C2123', `C2125', `C3111', `C3113', and `P493') and two cultivars (`Padrón' and `Yolo Wonder'). The percentage of cotyledon sections with shoot primordia after the induction phase was not genotype-dependent and always higher than with hypocotil sections (93.4% and 17.9%, respectively). The number of shoot primordia per responsive cotyledon section was also higher than with hypocotil sections (3.3 and 1.7, respectively). The genotype had a significant effect on the number of shoots regenerated per responsive cotyledon (1.1 to 5.5) or hypocotil (0.5 to 3.5) section. All adventitiously regenerated plants were fertile. This adventitious shoot regeneration protocol is being used to obtain transgenic plants from sweet bell pepper genotypes.


HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 461E-461
Author(s):  
Winthrop B. Phippen ◽  
James E. Simon

A plant regeneration protocol was successfully developed for basil (O. basilicum L.). Explants from 1-month-old seedlings yielded the highest frequency of regeneration of shoots (37%) with an average number of 3.6 shoots per explant. Calli and shoot induction were initiated on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ) (4 mg/L) for ≈30 days. Shoot induction and development was achieved by refreshing the induction medium once after 14 days. The most morphogenetically responsive explants were basal leaf explants from the first fully expanded true leafs of greenhouse-grown basil seedlings. Developing shoots were then rooted on MS media in the dark without TDZ. Within 20 days, rooted plantlets were transferred and acclimatized under greenhouse conditions where they developed normal morphological characteristics. This is the first report of a successful in vitro regeneration system for basil through primary callus. The establishment of a reliable regeneration procedure is critical when developing a transformation protocol for enhancing the production of basil for insect and disease resistance and improved essential oil constituents.


HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2129-2133 ◽  
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Min Deng ◽  
Jianjun Chen ◽  
Richard J. Henny

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.


HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Ana Carolina Espinosa ◽  
Paula M. Pijut ◽  
Charles H. Michler

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.


HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 813A-813
Author(s):  
Maria Cantor* ◽  
Rodica Pop ◽  
Ioana Pop

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.


HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1373-1377 ◽  
Author(s):  
Margarita Fraga ◽  
Mertxe Alonso ◽  
Marisé Borja

Meristem culture and/or thermotherapy were used for virus elimination from ornamental Phlox paniculata L. (`Blue Boy', `Orange perfection' and `Starfire') mother plants. Shoot tip, leaf, node and flower ovary explants collected from greenhouse-maintained virus free plants were cultured in vitro for shoot initiation. Adventitious shoot initiation was observed on Murashige and Skoog (MS) medium containing the cytokinin BA with or without the auxin NAA. The addition of 0.4 mg·L-1 thiamine, 0.4 mg·L-1 folic acid, and 40 mg·L-1 adenine sulfate to the MS medium did not improve the regeneration rate. Multiplication and rooting were genotype dependent. Blue Boy and Orange Perfection cultivars regenerated the maximum number of shoots from leaf explants. `Blue Boy' leaf explants from in vitro plants had a lower regeneration rate than explants from greenhouse plants. Cultivar `Starfire' had the highest shoot formation with open flower ovary explants and failed to regenerate from leaf explants. In vitro rooting of adventitious shoots in the presence of auxins (IAA, NAA, or IBA) with or without BA was less effective than ex vitro rooting. Chemical names used: 6-benzyladenine (BA); indole-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 696b-696
Author(s):  
Richard L. Bell

Discs of cambial tissue were excised from actively growing shoots of `Bartlett' pear, and explanted directly on regeneration induction media. The basal medium was 1/2 strength MS macro-nutrients, MS micro-nutrients and organics, 8 g/l agar, and 30 g/l sucrose. Phytohormone treatments consisted of a factorial design of NAA (0 and 5μM) and TDZ (1, 2, 3, 4, and 5μM). After 4 weeks incubation in the dark, the explants were transferred to auxin-free media with identical concentrations of TDZ. There was an absolute requirement for auxin in the induction medium, as all discs on auxin-free initial media died without callusing. Maximum shoot regeneration 4 weeks after transfer to expression media was obtained with an initial medium containing 5μM NAA and 3μM TDZ, from which 30% of the explants produced one or more adventitious shoots. This rate of regeneration is similar to that obtained in some experiments with in vitro leaf explants, and provides an alternative system for regeneration of pear.


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