Microspore embryogenesis in anther culture of three species of Populus and regeneration of dihaploid plants of Populustrichocarpa

1993 ◽  
Vol 23 (9) ◽  
pp. 1821-1825 ◽  
Author(s):  
Snorri Baldursson ◽  
Peter Krogstrup ◽  
Jens Viktor Nørgaard ◽  
Sven Bode Andersen

Microspore embryogenesis was induced from in vitro cultured anthers of Populusbalsamifera L., Populusmaximowiczii A. Henry, and Populustrichocarpa Torr. & Gray. Embryoids were formed at low frequencies on a modified Murashige and Skoog's medium, supplemented with 5 μM 6-benzylaminopurine, 5.1 mM L-glutamine, and 6% maltose. Growth regulator combinations (0–10 μM 6-benzylaminopurine and naphthaleneacetic acid) affected embryogenesis only slightly but formation of nonembryogenic callus from the anthers increased with increasing concentration of naphthaleneacetic acid. One donor clone of P. trichocarpa produced 54 embryoids from as many anthers during the two years of study. Adventitious shoots were obtained from 31 of these embryoids on woody plant medium with 2.5 μM 6-benzylaminopurine and 0.005 μM naphthaleneacetic acid. Adventitious shoots from 25 different embryoids were successfully rooted on woody plant medium containing 0.25 μM indole-3-butyric acid and transplanted to soil. Isozyme analysis confirmed microspore origin of all plants studied, and chromosome counts revealed that most of them had doubled their chromosome number spontaneously.

1994 ◽  
Vol 72 (2) ◽  
pp. 261-267 ◽  
Author(s):  
Conceição Eneida Silveira ◽  
Alain Cottignies

Propagation by stem cuttings and in vitro culture of apical bud explants were studied on Fraxinus excelsior L. Stem cuttings from 4- to 7-year-old trees growing under natural conditions sprouted only when cuttings were taken from dormant material. Only 6% of those that had sprouted developed roots by the 7th month of culture. Similarly, only apical bud explants harvested during the dormant period sprouted in vitro. Up to 87% of these sprouts developed two to four branching adventitious roots after 5 months of culture. During the initial phase of in vitro culture, the Quoirin and Lepoivre medium and the woody plant medium favoured sprout lengthening. During the phase of multiplication, up to three sprouts per explant developed with the woody plant medium in the presence of a combination of high 6-benzylaminopurine (3.0–4.0 mg∙L−1) and low indole-3-butyric acid (0.01–0.03 mg∙L−1) concentrations. Rooting was obtained in a medium without any growth regulators. Microscopic analysis showed a direct connection between the vascular elements of adventitious roots and stem of plantlet. Chromosome number in root apices of ash plantlets and ash trees grown under natural conditions was 2n = 46. Key words: chromosome number, Fraxinus excelsior L., in vitro plants, micropropagation, stem cuttings.


1995 ◽  
Vol 4 (5-6) ◽  
pp. 503-512
Author(s):  
Yingmou Yao

A protocol for micropropagation of sea buckthorn was developed starting with shoot tips or meristems from plants up to 18 years old. Among the different media used, the best medium for both initiation and multiplication was the woody plant medium (WPM). 6-Benzylaminopurine (BAP) was the most suitable growth regulator with an optimal concentration of 0.10-0.25 mg/l for initiation and 0.4-1.0 mg/l for multiplication. On WPM medium with BAP, the average rate of multiplication in Erlenmeyer flasks was 3.3-4.0 shoots per explant per month and in test tubes 2.0-3.0 shoots. Moreover, most explants produced several to tens of adventitious buds which grew into shoots. Explants rooted spontaneously in the multiplication medium at a frequency of about 33%. With this method, explants of different origins have been successfully propagated in vitro; and rooted young plants which had developed root nodules were produced both in the greenhouse and in the field.


2000 ◽  
Vol 48 (2) ◽  
pp. 215 ◽  
Author(s):  
J. Anthony ◽  
C. B. McLean ◽  
A. C. Lawrie

A system of micropropagation has been developed for Epacris impressa Labill. (pink heath) (Epacridaceae), the floral emblem of Victoria, Australia. Only explants from glasshouse-grown plants treated with 1.2 g L–1 mancozeb were established successfully in vitro. Shoot material was very sensitive to surface-sterilisation, with 0.5% NaOCl for 5 min being optimal. Multiple shooting was induced optimally on Woody Plant Medium (WPM, Lloyd and McCown 1980) with 12–25 µM of the cytokinin 2iP (6-(γ,γ-dimethylallylamino) purine). Inclusion of the auxin IBA (indole-3-butyric acid) induced callus and reduced shooting. Rooting in vitro was greatest (up to 40%) with half-strength WPM and 16 µM IBA. Clones from individual plants varied in multiple shooting response to 2iP (0–49 µM) and root induction response to auxins (IBA and NAA (α-naphthaleneacetic acid), 0–43 µM). These results suggest that explant materials are the main determinant of success in in vitro propagation and that they require individual optimisation of treatments to maximise shoot and root formation.


HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1052B-1052
Author(s):  
Denita Hadziabdic ◽  
Robert N. Trigiano ◽  
Stephen Garton ◽  
Mark T. Windham

Axillary and apical buds from five Cornus kousa cultivars (`Little Beauty', `Samaritan', `Heart Throb', `Rosabella', and `Christian Prince') were initially established on two basal media, woody plant medium (WPM) and woody plant medium/broad leaved tree medium (BW), amended with the following concentrations of 6–benzylaminopurine (BA): 0, 2, 4, and 8 μm. After explants were transferred at 4-week intervals for 28 weeks beginning in April, only microshoots of `Samaritan', `HeartThrob' and `Rosabella', were harvested from proliferating cultures and placed on rooting media. `Little Beauty' and `Christian Prince' did not perform well in multiplication phase of tissue culture and were excluded from further studies. Rooting media contained WPM or BW supplemented with either 1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA) or indole-3-acetic acid (IAA) at the following concentrations: 0, 0.5, 1.5, 4.5, and 13.5 μm. Six weeks following rooting experiment, preliminary data were collected and results showed that total of nine plants rooted on both WPM and BW media supplemented with IBA, 17 plants rooted on media supplemented with NAA, and 14 plants rooted on media supplemented with IAA. These results indicated that NAA and IAA appeared to be better for root production on C. kousa cultivar microshoots than IBA. Additionally, WPM supported more root production, when compared to BW, even though both media resulted in rooted microshoots. Proliferating masses were placed on fresh medium with 2μm BA and were used again for the rooting projects.


2018 ◽  
Vol 8 (3) ◽  
pp. 208-214
Author(s):  
Arum Sekar Wulandari ◽  
Erina Sulistiani ◽  
Esthi Liani Agustiani

Saninten (Castanopsis argentea (Blume) A.DC.) is one of the member Fagaceae family which can produce wood and non-wood product. Micropropagation of saninten by in vitro has never been reported. This research aims to identify the growth response of saninten shoot by adding Plant Growth Regulator (PGR), they are 6-benzylaminopurine (BAP) and indole-3-acetic-acid (IAA) with different concentrations in propagation in vitro on woody plant medium (WPM). The research conducted a completely randomized design (CRD) factorial. They are PGR type and PGR concentration. The PGR type consist of two levels namely BAP and IAA. PGR concentration consist of three levels namely 0 mg/L BAP and IAA, 0.5 mg/L BAP or 0.1 mg/L IAA, and 1.0 mg/L BAP or 0.1 mg/L IAA. Parameters observed is the amount of shoot, shoot height, and callus growth. The combination of BAP (0, 0.5, 1 mg/L) and IAA (0, 0.1, 0.1 mg/L) haven’t produce optimal growth of shoot. WPM medium with 0.5 mg/L BAP was able to produce the best of percentage of shoot, the number of shoots, and shoot height growth. WPM medium with IAA concentration of 0.1 and 0.2 mg/L produce explant with callus.Key words: 6-benzylaminopurine (BAP), Castanopsis argentea, indole-3-acetic-acid (IAA), woody plant medium (WPM)


2007 ◽  
Vol 8 (1) ◽  
pp. 22 ◽  
Author(s):  
Fabiola Ocampo ◽  
Víctor Manuel Núñez

<p>Se indujeron múltiples brotes mediante organogénesis directa a partir de segmentos nodales de 10 genotipos diferentes de guayaba. Para ello se estableció un sistema de propagación clonal <em>in vitro </em>combinado con inducción rápida de brotes <em>ex vitro </em>para propagar árboles élite. La utilización de segmentos nodales permitió obtener en poco tiempo brotes adventicios adecuados para multiplicación masiva. La respuesta <em>in vitro </em>de los genotipos fue evaluada usando los medios de cultivo MS (Murashige y Skoog, 1962), Mc (Mascarenhas) y WPM (<em>Woody Plant Medium</em>) suplementados con 0,1 mg•L-1 de ácido indolácetico (AIA) y 0,25 mg•L<sup>-1</sup> de bencilaminopurina (BAP). El procedimiento de desinfección con hipoclorito de sodio previno eficientemente la contaminación de los explantes después de la inoculación en el medio de cultivo. El mayor porcentaje en la inducción de brotes se logró con 0,25 mg•L<sup>-1</sup> de BAP. Los segmentos nodales presentaron de 1 a 2 brotes adventicios por explante después de 15 días de inoculados y de 3 a 7 brotes a los 30 días después del inicio del cultivo. Una vez individualizados los brotes se usaron en una nueva fase de multiplicación masiva en la que se probaron cuatro sustratos diferentes durante el enraizamiento y el endurecimiento. Esta metodología permitió la propagación <em>in vitro </em>de guayaba cuatro semanas después del inicio del cultivo. Los mejores resultados se lograron con el medio WPM que permitió obtener las primeras plántulas enraizadas dos semanas después de la transferencia al sustrato de enraizamiento.</p><p> </p><p><strong>In vitro propagation of Psidium guajaba using direct organogenesis from nodal segments</strong></p><p>Multiple shoots were induced using direct organogenesis from nodal segments of 10 different genotypes of guayaba. For this an in vitro clonal propagation system combined with rapid ex vitro induction of shoots was established in order to propagate elite trees. The use of nodal segments resulted in adventitious shoots adequate for mass multiplication in less time. The in vitro response of the genotypes was evaluated using the culture media MS (Murashige and Skoog, 1962), Mc (Mascarenhas) and WPM (Woody Plant Medium) supplemented with 0.1 mg·L-1 of indole acetic acid (IAA) and 0.25 mg·L-1 of benzoaminopurine (BAP). The procedure for sterilizing with sodium hypochlorite effectively prevented the contamination of the explants after the inoculation of the culture medium. The greatest percentage of shoot induction was achieved with 0.25 mg·L-1 of BAP. The nodal segments showed between 1-2 adventitious shoots per explant 15 days post-inoculation and 3-7 shoots 30 days post-inoculation. Once individualized, the shoots were used in a new mass multiplication phase in which four different substrates were tested during rooting and hardening. This methodology permitted the in vitro propagation of guayaba four weeks post-inoculation. The best results were achieved with the WPM medium that resulted in the first rooted plantlets two weeks after the transfer to the rooting substrate.</p>


HortScience ◽  
1997 ◽  
Vol 32 (2) ◽  
pp. 312-314 ◽  
Author(s):  
John L. Edson ◽  
David L. Wenny ◽  
Annette Leege-Brusven

In vitro—derived microshoots of antelope bitterbrush, incubated for 1 month in media supplemented with 0.44 μm BA, grew 0.8 and 1.1 cm longer in woody plant medium (WPM) compared to full-strength and half-strength Murashige and Skoog (MS) media, respectively. Explants cultured in WPM supplemented with 0.44 μm BA and 0.54 μm NAA produced a mean of five axillary shoots per explant. Explants dipped in 0.1% IBA or 0.1% NAA rooted best in 0.1% IBA with 89% success ex vitro vs. 60% success in vitro. Survival of acclimatized plantlets rooted ex vitro was 95%, while 50% survived when rooted in vitro. After 1 year of greenhouse growth, 98% of plantlets survived and flowered. Chemical names used: benzyladenine (BA), 3-indolebutyric acid (IBA), 1-naphthaleneacetic acid (NAA).


HortScience ◽  
2018 ◽  
Vol 53 (7) ◽  
pp. 1045-1049 ◽  
Author(s):  
Dongliang Qiu ◽  
Xiangying Wei ◽  
Shufang Fan ◽  
Dawei Jian ◽  
Jianjun Chen

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.


HortScience ◽  
1991 ◽  
Vol 26 (12) ◽  
pp. 1555-1557
Author(s):  
Thomas W. Zimmerman ◽  
Fred T. Davies ◽  
Jayne M. Zajicek

Dyssodia pentacheta, a prostrate-growing perennial Texas wildflower with potential for use in low-maintenance landscapes, was propagated in vitro and by stem cuttings under mist. Optimum rooting for IBA-treated semihardwood terminal stem cuttings (3 to 30 mm IBA) and in vitro-grown nodal segments (30 to 100 mm IBA) occurred after 4 weeks under an intermittent mist system. A 300-mm IBA basal dip was lethal to macroand microcuttings. In vitro, D. pentacheta produced more shoots per nodal explant on Woody Plant Medium (2 g Gelrite/liter) with 1 to 10 μ m BA than with combinations of BA and 0.5 μm NAA. After shoot proliferation, the shoots were subculture twice and grown on growth regulator-free medium. When maintaining D. pentacheta in vitro on media devoid of plant growth regulators, 1% sucrose was more effective than 2% for promoting shoot growth and suppressing apparent production of phenolics. Chemical names used: N-(phenylmethyl) -1H-purin-6-amine (BA); 1H-indole-3-butyric acid (IBA); 1-naphthaleneacetic acid (NAA).


2012 ◽  
Vol 39 (No. 1) ◽  
pp. 21-25 ◽  
Author(s):  
J. Sedlák ◽  
F. Paprštein

The goal of this study was to investigate in vitro multiplication protocols for use with red currant cultivars grown in the Czech Republic. Cultivars Detvan, Vitan and Rotte H&ouml;llandische were successfully established in vitro using mercuric chloride in a concentration of 0.15% as a sterilization solution. The overall rate of contamination was 25.7%. Two proliferation media Murashige and Skoog medium (MS) and McCown woody plant medium (WPM) containing 1 or 2&nbsp;mg/l of 6-benzylaminopurine (BAP) were tested. Initial explants produced new plants in the form of rosettes. Rosettes arose from the base of the initial explants in the form of adventitious bud formation. The shoot number was relatively low and varied between 1.0 and 2.1. Generally, the highest number was obtained for cultivar Rotte Holl&auml;ndische that produced 2.1 &plusmn; 0.1 new rosettes on MS medium containing lower concentration 1 mg/l BAP. In contrary, Vitan cv. had significantly lower shoot number ranging from 1.0 to 1.3. WPM medium with a lower concentration of mineral salts proved to be unsuitable for the multiplication of tested cultivars.


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