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

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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SHOOT ORGANOGENESIS FROM CAMBIAL DISC EXPLANTS OF PEAR.

HortScience ◽

10.21273/hortsci.27.6.696b ◽

1992 ◽

Vol 27 (6) ◽

pp. 696b-696

Author(s):

Richard L. Bell

Keyword(s):

Shoot Organogenesis ◽

Adventitious Shoots ◽

Leaf Explants ◽

Basal Medium ◽

Induction Medium ◽

Absolute Requirement ◽

Initial Medium ◽

Free Media ◽

Macro Nutrients

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

HortScience ◽

10.21273/hortsci.42.3.721 ◽

2007 ◽

Vol 42 (3) ◽

pp. 721-723 ◽

Cited By ~ 6

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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In vitro propagation of Crambe maritima

Canadian Journal of Botany ◽

10.1139/b87-010 ◽

1987 ◽

Vol 65 (1) ◽

pp. 72-75 ◽

Cited By ~ 2

Author(s):

J. Y. Peron ◽

E. Regnier

Keyword(s):

In Vitro Propagation ◽

Indoleacetic Acid ◽

Adventitious Shoots ◽

Basal Medium ◽

Naphthaleneacetic Acid ◽

Petiole Explants ◽

Acclimatization Period

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.

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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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117 An In Vitro Regeneration System for Basil (Ocimum basilicum L.)

HortScience ◽

10.21273/hortsci.34.3.461e ◽

1999 ◽

Vol 34 (3) ◽

pp. 461E-461

Author(s):

Winthrop B. Phippen ◽

James E. Simon

Keyword(s):

In Vitro Regeneration ◽

Morphological Characteristics ◽

Leaf Explants ◽

Basal Medium ◽

Induction Medium ◽

Shoot Induction ◽

Regeneration System ◽

Transformation Protocol ◽

Regeneration Procedure

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.

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Regeneration and Genetic Transformation of Arabidopsis Pumila: An Ephemeral Plant Suitable for Investigating the Mechanisms for Adaptation to Desert Environments

10.21203/rs.3.rs-160586/v1 ◽

2021 ◽

Author(s):

Yuhuan Jin ◽

Li Guo ◽

Danqing Liu ◽

Yongguang Li ◽

Hao Ai ◽

...

Keyword(s):

Adventitious Shoots ◽

Cellular Level ◽

Cauliflower Mosaic Virus ◽

Induction Medium ◽

35S Promoter ◽

Naphthalene Acetic Acid ◽

Growth Duration ◽

Petiole Explants ◽

Induction Rate ◽

Ephemeral Plant

Abstract Arabidopsis pumila is a type of cruciferous ephemeral plant, which in China mainly grows in the desert environments of northern Xinjiang. A. pumila not only has a short growth duration, but also has high photosynthetic efficiency, seed yield, salt tolerance, and drought resistance. It is an ideal species for the study of environmental adaptations in ephemeral plants. We induced callus tissue formation on the roots and hypocotyls of 8-day-old seedlings, and on the leaves and petioles of 4-week-old seedlings, and obtained multiple adventitious shoots on these tissues grown on Murashige and Skoog induction medium supplemented with 0.5 mg/L 6-Benzylaminopurine and 0.1 mg/L α-Naphthalene acetic acid. Young roots, hypocotyls, leaves, and petioles could all induce calluses, but the induction rate was highest on young roots. In addition, the leaves and petioles of 4-week-old seedlings were used as explants, the Δ1-pyrroline-5-carboxylic acid synthase gene 1 of A. pumila controlled by 35S promoter of cauliflower mosaic virus was used as target gene, and hygromycin B was used as screening antibiotic to explore Agrobacterium tumefaciens GV3101 mediated transformation. The results showed that the callus induction rate of petiole explants was the highest when they were treated with Agrobacterium suspension (OD600 = 0.6) for 10 min and thenco-cultured in dark for 2 d. The qRT-PCR results showed that the ApP5CS1.1 gene was overexpressed in the transgenic plants. These protocols provide working research methods for exploring the cellular level adaptative mechanisms of this species to desert environments.

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In vitro Regeneration of Grass Pea (Lathyrus sativus L.)

Jahangirnagar University Journal of Biological Sciences ◽

10.3329/jujbs.v4i2.27790 ◽

2016 ◽

Vol 4 (2) ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

P Saha ◽

M Afrin ◽

AKM Mohiuddin ◽

AM Shohael

Keyword(s):

In Vitro Regeneration ◽

Basal Medium ◽

Multiple Shoots ◽

Nodal Explants ◽

Lathyrus Sativus ◽

Naphthalene Acetic Acid ◽

Grass Pea ◽

Half Strength ◽

Lathyrus Sativus L

In vitro regeneration protocol for grass pea (Lathyrus sativus L.) was optimized using different concentrations and combinations of growth regulators. Direct shoot regeneration obtained through shoot organogenesis from different explants of grass pea cultured on MS medium supplemented with Gamborg B5 vitamin containing 6-benzylaminopurine (BAP), Thidiazuron (TDZ) and ?-naphthalene acetic acid (NAA). Highest percentage of shoots were obtained at 4.0 mg/l of BAP on nodal explants. Stunted multiple shoots were developed from nodal explants while 1.5 mg/l TDZ was used. About 56% of direct shoots were also obtained, while the combination of BAP (4.0 mg/l) and NAA (0.5 mg/l) were used. Regenerated plantlets were rooted most effectively (40%) in rooting medium containing half strength of MS basal medium containing 1.0 mg/l NAA. Well rooted plantlets were further successfully acclimatized to ambient humidity level and grown in controlled environment until hardening.Jahangirnagar University J. Biol. Sci. 4(2): 1-8, 2015 (December)

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Regeneration of Blueberry Cultivars through Indirect Shoot Organogenesis

HortScience ◽

10.21273/hortsci13059-18 ◽

2018 ◽

Vol 53 (7) ◽

pp. 1045-1049 ◽

Cited By ~ 3

Author(s):

Dongliang Qiu ◽

Xiangying Wei ◽

Shufang Fan ◽

Dawei Jian ◽

Jianjun Chen

Keyword(s):

Genetic Background ◽

Woody Plant ◽

Shoot Organogenesis ◽

Adventitious Shoots ◽

Leaf Explants ◽

Ex Vitro ◽

Significant Difference ◽

Woody Plant Medium ◽

Cultivar Improvement

Leaf explants derived from in vitro–grown shoots of blueberry cultivars Bluejay, Pink Lemonade, Sunshine Blue, and Top Hat were cultured on woody plant medium (WPM) supplemented with 9.12 μm 6-(4-hydroxy-3-methylbut-2-enylamino) purine or zeatin (ZT) in combination with 1.23, 2.46, or 4.92 μm indole-3-butyric acid (IBA). Calluses were induced from the explants and adventitious shoots were regenerated. ‘Sunshine Blue’ and ‘Top Hat’ produced more than four shoots per explant but shoot numbers were less than one for each ‘Pink Lemonade’ explant and about 0.2 per ‘Bluejay’ explant. The results indicate that there is significant difference among cultivars in indirect shoot organogenesis. The differences may be related to their diverse genetic background as they are polyploid hybrids. Microcuttings derived from adventitious shoots of ‘Sunshine Blue’ rooted in vitro in WPM medium supplemented with 9.84 μm IBA and also rooted ex vitro in a peat-based substrate after cuttings were dipped or not dipped in IBA solutions. Direct rooting of microcuttings in the peat-based substrate was effective, suggesting that in vitro rooting may not be necessarily needed. Survival rate of ex vitro–rooted plants in a shaded greenhouse was high, more than 90%. The established shoot regeneration protocols could be used for rapid propagation of ‘Sunshine Blue’ and ‘Top Hat’ and for cultivar improvement through genetic transformation.

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