scholarly journals In vitro Propagation and Assessment of Genetic Relationships of Citrus Rootstocks Using ISSR Molecular Markers

2017 ◽  
Vol 45 (2) ◽  
pp. 383-391 ◽  
Author(s):  
Constantinos SALIS ◽  
Ioannis E. PAPADAKIS ◽  
Spyridon KINTZIOS ◽  
Marianna HAGIDIMITRIOU
Keyword(s):  
Molecular Markers ◽  
Genetic Variability ◽  
Root Length ◽  
In Vitro Propagation ◽  
Genetic Relationships ◽  
Shoot Proliferation ◽  
Poncirus Trifoliata ◽  
Naphthaleneacetic Acid ◽  
Issr Molecular Markers

The behavior of six citrus rootstocks, Volkameriana, Citrumelo ‘Swingle’, Citrange ‘Carrizo’, Poncirus trifoliata ‘Serra’, Poncirus trifoliata ‘Rubidoux’ and Poncirus trifoliata ‘Flying Dragon’, in in vitro propagation was studied and compared for shoot proliferation and rooting. In addition, the genetic relationships among the rootstocks studied and other Citrus species, using the Inter-Simple Sequence Repeats (ISSR) molecular markers, were investigated. Nodal explants of three months old shoots were used in Murashige and Skoog medium supplemented with N6-benzyladenine (BA) for shoot proliferation and with naphthaleneacetic acid (NAA) for rooting. The rootstock Volkameriana showed a statistically significant higher number of shoots (1.81), shoot length (15.14 mm) and number of leaves per explant (5.81), while all three Poncirus trifoliata rootstocks showed the lowest numbers. The number of roots and root length per explant were evaluated at the end of the rooting phase. The rootstock ‘Swingle’ showed a higher number of roots per explant (4.2) followed by ‘Flying Dragon’ (3.93) and ‘Carrizo’ (3.23) rootstocks. The rootstocks ‘Swingle’ (140.8 mm), Volkameriana (148 mm) and ‘Flying Dragon’ (131.12 mm) had significantly higher root length per explant compared to ‘Carrizo’ (31 mm) and ‘Rubidoux’ (34.5 mm). The ISSR molecular marker technique used in the present study grouped successfully the different species, varieties and rootstocks studied, revealing their genetic variability. The genetic variability observed among the rootstocks ranged between 0.29 (Poncirus trifoliata ‘Serra’ and Citrumelo ‘Swingle’) and 0.60 (Volkameriana and Citrumelo ‘Swingle’). The response of the rootstocks studied in in vitro propagation however is not related to their genetic affinity.

scholarly journals In Vitro Propagation of Eriostemon myoporoides and Eriostemon `Stardust'

HortScience ◽  
1994 ◽  
Vol 29 (6) ◽  
pp. 686-688 ◽  
Author(s):  
James R. Ault
Keyword(s):  
Root Length ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Length ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Axillary Shoot Proliferation

Optimal axillary shoot proliferation was obtained from stem explants of a clone of Eriostemon myoporoides DC. on Murashige and Skoog (MS) basal medium containing 0.1 mg BA/liter, and of Eriostemon `Stardust' on MS medium containing 0.5 mg BA/liter. Overall average number of shoots and shoot lengths for all treatments was greater for E. `Stardust' (22.4 shoots and 12.1-mm shoot length) than for E. myoporoides (4.5 shoots and 8.3-mm shoot length). Maximum percent rooting of E. myoporoides (42%) and E. `Stardust' (95%) was obtained on MS medium supplemented with 1.0 mg K-IBA/liter for E. myoporoides and 0.1 mg NAA/liter for E. `Stardust'. Overall average percent rooting and root lengths were greater for E. `Stardust' (42% rooting and 11.0-mm root length) than for E. myoporoides (27% rooting and 2.3-mm root length). For E. `Stardust', reducing sucrose in the rooting medium from 50 to 25 g·liter-1 significantly decreased overall average percent rooting to 1670 and root length to 6.8 mm. Plantlets of both clones were acclimatized in the greenhouse and transferred successfully to soil, although survival was <7070. Chemical names used: N -(phenylmethyl) -l H -purine-6-amine (BA); potassium-l H -indole-3-butyric acid (K-IBA); l-naphthaleneacetic acid (NAA).

scholarly journals Plant Growth Regulator Effects on In Vitro Propagation and Stevioside Production in Stevia rebaudiana Bertoni

HortScience ◽  
2016 ◽  
Vol 51 (12) ◽  
pp. 1573-1580 ◽  
Author(s):  
Bärbel Röck-Okuyucu ◽  
Meltem Bayraktar ◽  
Ismail Hakki Akgun ◽  
Aynur Gurel
Keyword(s):  
Plant Growth ◽  
In Vitro Propagation ◽  
Shoot Growth ◽  
Morphological Changes ◽  
Stevia Rebaudiana ◽  
Shoot Proliferation ◽  
Naphthaleneacetic Acid ◽  
Steviol Glycosides ◽  
Control Medium

Stevia rebaudiana is of great importance due to its steviol glycosides (SGs) which are natural sweeteners used by the food industry as well as having medicinal purposes. In the present study, the effect of plant growth regulators (PGRs) and explant types on in vitro propagation and shoot growth of S. rebaudiana were studied, the effect of PGRs on SGs production was determined. For this purpose, nodal explants and shoot tip explants were cultured on woody plant medium (WPM) supplemented with cytokinins [6-benzyladenine (BA), kinetin (Kn), or thidiazuron (TDZ)] or cytokinins + auxins combinations [BA + indoleacetic acid (IAA); BA + naphthaleneacetic acid (NAA); Kn + IAA; Kn + NAA]. Although, the best shoot proliferation was obtained on WPM supplemented with BA + NAA combinations, shoots grown on PGR-containing media produced callus at the base of the shoots and showed chlorosis and necrosis. Additionally, shoots showed at all concentrations of TDZ, and at higher concentrations of BA, morphological changes such as malformed leaves and poor shoot growth. In contrast to PGR-containing media, on the PGR-free control medium, the development of shoots and roots occurred simultaneously and healthy and well-developed plantlets were obtained. Thus, we developed an economical viable means of in vitro propagation by minimizing the micropropagation steps and removing the requirement of PGRs. According to the high-performance liquid chromatography (HPLC) results, PGR-free control medium (WPM) led to considerably higher stevioside content in the leaves compared with the PGR(s)-containing media and the highest stevioside [34 mg·g−1 dry weight (DW)] and rebaudioside A content was only detected on the control medium without PGRs. Steviolbioside, rubusoside, and dulcoside A were detected qualitatively in the leaves of shoots grown on WPM supplemented with 2.27 μm TDZ, 4.54 μm TDZ, 2.22 μm BA + 2.69 μm NAA, 2.22 μm BA + 5.37 μm NAA, 2.32 μm Kn + 5.71 μm IAA, or 2.32 μm Kn + 2.69 μm NAA.

scholarly journals In Vitro Propagation of Eucomis autumnalis, E. comosa, and E. zambesiaca by Twin-scaling

HortScience ◽  
1995 ◽  
Vol 30 (7) ◽  
pp. 1441-1442 ◽  
Author(s):  
James R. Ault
Keyword(s):  
In Vitro Propagation ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Plant Survival ◽  
Single Shoot

Shoot formed in vitro from twin-scale explants of Eucomis autumnalis (Mill.) Chitt., E. comosa (Houtt.) Wehrh., and E. zambesiaca Bak. cultured on Murashige and Skoog (MS) basal medium containing 0.0, 4.4, 11.1, or 22.2 μm BA and 0.0 or 5.4 μm NAA. In all three species, shoot proliferation was obtained from single-shoot explants subcultured on medium supplemented with 4.4, 11.1, or 22.2 μm BA and 0.0 or 5.4 μm NAA. Shoots of all three species rooted readily on MS medium supplemented with 0.0, 2.7, 5.4, or 10.8 mm NAA. Overall rooting percentages were 95%, 98%, and 100% for E. autumnalis, E. comosa, and E. zambesiaca, respectively. Plant survival for rooted shoots of all three species was 100% following transfer to a 1 perlite: 1 peat (v/v) medium in the greenhouse. Chemical names used: 6-benzyladenine (BA); 1-naphthaleneacetic acid (NAA).

scholarly journals Cytokinin-Facilitated Plant Regeneration of Three Brachystelma Species with Different Conservation Status

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1657
Author(s):  
Nqobile P. Hlophe ◽  
Adeyemi O. Aremu ◽  
Karel Doležal ◽  
Johannes Van Staden ◽  
Jeffrey F. Finnie
Keyword(s):  
In Vitro Propagation ◽  
Slow Growth ◽  
Conservation Status ◽  
Shoot Proliferation ◽  
Nodal Segments ◽  
Valuable Resource ◽  
Ex Vitro ◽  
Nutritional Values ◽  
Over Exploitation

In Africa and Asia, members of the genus Brachystelma are well-known for their diverse uses, especially their medicinal and nutritional values. However, the use of many Brachystelma species as a valuable resource is generally accompanied by the concern of over-exploitation attributed to their slow growth and general small size. The aim of the current study was to establish efficient micropropagation protocols for three Brachystelma species, namely Brachystelma ngomense (endangered), Brachystelma pulchellum (vulnerable) and Brachystelma pygmaeum (least concern), as a means of ensuring their conservation and survival. This was achieved using nodal segments (~10 mm in length) as the source of explants in the presence of different concentrations of three cytokinins (CK) namely N6-benzyladenine (BA), isopentenyladenine (iP) and meta-topolin riboside (mTR), over a period of 6 weeks. The highest (25 µM) concentration of cytokinin treatments typically resulted in significantly higher shoot proliferation. However, each species differed in its response to specific CK: the optimal concentrations were 25 µM mTR, 25 µM iP and 25 µM BA for Brachystelma ngomense, Brachystelma pulchellum and Brachystelma pygmaeum, respectively. During the in vitro propagation, both Brachystelma ngomense and Brachystelma pygmaeum rooted poorly while regenerated Brachystelma pulchellum generally lacked roots regardless of the CK treatments. Following pulsing (dipping) treatment of in vitro-regenerated shoots with indole-3-butyric acid (IBA), acclimatization of all three Brachystelma species remained extremely limited due to poor rooting ex vitro. To the best of our knowledge, the current protocols provide the first successful report for these Brachystelma species. However, further research remains essential to enhance the efficiency of the devised protocol.

In vitro propagation of Crambe maritima

1987 ◽  
Vol 65 (1) ◽  
pp. 72-75 ◽  
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.

scholarly journals Micropropagation of Clerodendrum phlomidis L.F.

2016 ◽  
Vol 24 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Mafatlal M. Kher ◽  
Deepak Soner ◽  
Neha Srivastava ◽  
Murugan Nataraj ◽  
Jaime A. Teixeira da Silva
Keyword(s):  
Callus Formation ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Explants ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoot Proliferation ◽  
Clerodendrum Phlomidis

Abstract Clerodendrum phlomidis L. f. is an important medicinal plant of the Lamiaceae family, particularly its roots, which are used for various therapeutic purposes in a pulverized form. The objective of this study was to develop a standard protocol for axillary shoot proliferation and rooting of C. phlomidis for its propagation and conservation. Nodal explants were inoculated on Murashige and Skoog (MS) medium that was supplemented with one of six cytokinins: 6-benzyladenine, kinetin, thidiazuron, N6-(2-isopentenyl) adenine (2iP), trans-zeatin (Zea) and meta-topolin. Callus induction, which was prolific at all concentrations, formed at the base of nodal explants and hindered shoot multiplication and elongation. To avoid or reduce callus formation with the objective of increasing shoot formation, the same six cytokinins were combined with 4 μM 2,3,5-tri-iodobenzoic acid (TIBA) alone or in combination with 270 μM adenine sulphate (AdS). Nodal explants that were cultured on the medium supplemented with 9.12 μM Zea, 4 μM TIBA and 270 μM AdS produced significantly more and longer shoots than on medium without TIBA and AdS. Half-strength MS medium supplemented with 8.05 μM α-naphthaleneacetic acid was the best medium for root formation. Most (75%) in vitro rooted plantlets were successfully acclimatized under natural conditions.

scholarly journals In Vitro Propagation of Agave americana by Indirect Organogenesis

HortScience ◽  
2017 ◽  
Vol 52 (7) ◽  
pp. 996-999 ◽  
Author(s):  
Carlos Alberto Lecona-Guzmán ◽  
Sheila Reyes-Zambrano ◽  
Felipe Alonso Barredo-Pool ◽  
Miguel Abud-Archila ◽  
Joaquín Adolfo Montes-Molina ◽  
...  
Keyword(s):  
In Vitro Propagation ◽  
Shoot Proliferation ◽  
Ms Medium ◽  
Indolebutyric Acid ◽  
Indirect Organogenesis ◽  
Meristematic Tissue ◽  
Phenotypic Alteration ◽  
Sexual And Asexual Reproduction ◽  
Agave Americana

Factors such as slow growth, low rates of sexual and asexual reproduction, and viability of seeds among others limit the massive propagation of Agave americana L. by conventional methods. In this study, callus induction and shoot proliferation was determined in A. americana using Murashige and Skoog (MS) medium supplemented with dicholorophenoxyacetic acid (2,4-D) and 6-benzyl adenine (BA). Meristematic tissue was used as the explants, and were placed on MS medium supplemented with 30.0 g·L−1 sucrose with 0.11, 0.18, or 0.45 μm 2,4-D and 11.0, 22.0, 38.2, 44.0, 58.7, or 73.3 μm BA. Treatments were implemented according to factorial experimental design 3 × 6. After 1 month, the number of explants with callus was determined, whereas the numbers of shoots per explant were monitored after 4, 16, 20, and 36 weeks. The maximum percent of explants with callus was obtained with 0.11 μm 2,4-D and 58.7 and 73.3 μm BA, whereas the maximum numbers of shoots per explant (71) were obtained with 0.11 μm 2,4-D and 73.3 μm BA. The effect of different concentrations of indolebutyric acid (IBA) in the rooting of shoots was evaluated. There were no significant effects of IBA on the number of roots, root length, and axillary roots. Plantlets were acclimatized in the glasshouse and they did not show any phenotypic alteration. This is a highly efficient protocol for the in vitro propagation of A. americana via indirect organogenesis.

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