scholarly journals Micropropagation of Betula platyphylla ‘Fargo’ via Shoot Tip Culture and Regeneration from Leaf Tissues

2000 ◽  
Vol 18 (2) ◽  
pp. 119-122 ◽  
Author(s):  
Zong-Ming Cheng ◽  
Jeffrey P. Schnurr ◽  
Wenhao Dai

Abstract A micropropagation system was developed for mass propagation of ‘Fargo’® (Dakota Pinnacle™), a newly released cultivar of Asian white birch (Betula platyphylla). Shoot tips from the mature, 7-year-old tree were established on 75% strength Murashige and Skoog medium supplemented with 0.1 μM thidiazuron. The greatest shoot proliferation occurred on Woody Plant Medium supplemented with 2.2 μM benzyladenine (BA), solidified with 6.5 g/liter agar, and cultured at 24C (75F). Microshoots were rooted in vitro or ex vitro followed by establishment in the greenhouse. A system to regenerate plantlets from leaves of aseptically cultured shoots was also developed. The optimum conditions for shoot regeneration from leaves included a 2-week dark treatment before exposure to a 16/8 hour light/dark photoperiod, use of large and healthy leaf explants, and culture on the Woody Plant Medium containing 20.0 or 30.0 μM BA. The regenerated shoots proliferated on the micropropagation medium and were divided, and the resulting shoots were rooted ex vitro and acclimated in greenhouse conditions.

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 515e-515
Author(s):  
Jeffrey P. Schnurr ◽  
Zongming Cheng

A selection of Betula platyphylla, from an open pollinated population, was made for upright growth habit, cold hardiness, and a dark green canopy. A micropropagation system was developed to overcome the difficulty with conventional propagation techniques. Shoot-tip cultures were best established in 3/4 strength MS medium supplemented with 0.1 μM thiadiazuron. After 5 weeks in culture, shoots were transferred to woody plant medium (WPM) with 4.4 μM BA. The highest proliferation rate occurred at 24 C on WPM, solidified with agar, and supplemented with 2.2 μM BA. Shoots rooted in vitro and ex vitro and have been established in the field. A regeneration system has also been developed using leaves from aseptic cultures. The optimum conditions for shoot regeneration include a 2-week dark treatment before exposure to a 16-h day/8-h night cycle. Large, healthy leaf explants cultured on WPM with 20 μM BA regenerated shoots at the highest frequency. Regenerated shoots, when transferred to the micropropagation system, proliferate successfully. Currently, a transformation system for this selection is being developed.


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.


Author(s):  
Hieu Trung Tran ◽  
Chung Van Huynh ◽  
Hue Thi Linh Bui ◽  
Ngan Thi My Luong ◽  
Anh Lan Bui ◽  
...  

Paramignya trimera (Oliv.) Guill., a woody climber commonly known as "Xao tam phan", has been used in Vietnamese folk for the treatment of numerous cancers. Due to word of mouth about the anticancer properties of this plant, its stems and roots have been overexploited leading to the serious decline of this species in Phu Yen, Khanh Hoa and Ninh Thuan provinces. The aim of the study was to establish an in vitro propagation protocol for the conservation of P. trimera. In this research shoot clusters (5–8 shoots/cluster) were regenerated from axillary bud explants of 1–3 year-old trees after 3 months of cultures on the WPM (woody plant medium) supplemented with STS 3 and BA 5–7 mg/L. STS (silver thiosulfate) was used to prevent the leaf abscission. These shoot clusters grew slowly and reached 1–3 cm in heights after 4 months of the cultures. These shoot clusters did not form any roots after 2 months of culture on the rooting media with IBA and/or NAA 1–5 mg/L. However, there was 51 % of the treated shoot clusters acclimatized and produced new stem and leaves after 2 months growing in greenhouse. WPM supplemented with STS 3, BA 5 and IBA 5 mg/L showed the best response for callus induction in leaf explants after 3 months of cultures. Among the callus types, the milky white compact calli were induced at the cut surface of leaf explants after 3 months of the cultures and became the compact and nodulated calli within 4 weeks later.


HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1096a-1096
Author(s):  
Mark H. Brand ◽  
William G. Cullina

Increasing interest in landscape use of Aronia arbutifolia and A. melanocarpa has led to the establishment of breeding programs and selection of improved phenotypes within the genus. A rnicropropagation system was developed to facilitate rapid and easy multiplication of improved forms of Aronia. Actively growing shoot tips of A. arbutifolia `Brilliantissima' and A. melanocarpa were used to initiate shoot proliferation from axillary buds. Optimum proliferation of shoots useful for micropropagation occurred on media supplemented with 0.5 to 1.0 mg 1-1 benzyladenine. Both Murashige and Skoog medium and Woody Plant medium supported vigorous shoot proliferation, but differences in culture morphology were evident. In vitro rooting and non-sterile rooting methods both resulted in high rooting percentages, the formation of numerous roots and subsequent rapid growth of plantlets.


HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 757A-757 ◽  
Author(s):  
Guochen Yang ◽  
Paul E. Read ◽  
Marihelen Kamp-Glass

Chestnut (Castanea spp.) is considered difficult to micropropagate. The timing for harvesting explant materials from forced stems is critical, although many factors need to be considered for successful micropropagation. Previous research with spirea and five-leaf aralia demonstrated that forcing solution techniques extended the availability of high-quality explant material, thus expediting micropropagation. However, preliminary research illustrated that chestnut is very difficult to force and the new forced softwood growth is very short-lived, which made micropropagation difficult. It was found that, at about 7 days from budbreak, the forced chestnut softwood growth (about 2 cm long) served as the best explant material. If longer than this timing window, the new growth would die. If shorter, the explants had a high contamination rate, exudation of purported phenolic compounds, and explants would not regenerate. Shoot proliferation and callus regeneration were achieved by culturing good-quality explants on Woody Plant Medium supplemented with 0.1 mg BA/liter. The new shoots grew vigorously in vitro with apparent normal morphology.


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 ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 397C-397
Author(s):  
Guochen Yang ◽  
Marihelen Kamp-Glass

Exochorda racemosa is an ornamental shrub with white flowers that is spiraea-like, deciduous, and hardy. The buds resemble pearls. Normally it is propagated by seeds, layers, and cuttings of softwood. However, it is a slow process that takes a few years to produce a reasonable size plant for the demanding market. Our objective was to establish a successful in vitro culture and to rapidly multiply this ornamental species. Softwood explant materials were collected from a local nursery and were disinfested with 15% bleach solution and rinsed three times with sterile distilled and deionized water. In vitro cultures were established and maintained in woody plant medium (WPM) supplemented with BA at 0.1 mg·L-1, 3% sucrose, and 0.7% agar with the pH adjusted to 5.8. Then shoots were transferred to the multiplication medium containing BA, CPPU, or thidiazuron (TDZ) at various concentrations. Preliminary results show that explants cultured on medium containing TDZ produced the best shoot proliferation.


2008 ◽  
Author(s):  
Επαμεινώνδας Κάρτσωνας

Στην παρούσα εργασία αναπτύχθηκε μέθοδος μικροπολλαπλασιασμού του Quercus euboica Pap, ενός σπάνιου ενδημικού είδους βαλανιδιάς της Ελλάδας. Έκφυτα κόμβων από σπορόφυτα έδωσαν τον υψηλότερο ρυθμό πολλαπλασιασμού από αντίστοιχα ενήλικων φυτών. Ο υψηλότερος ρυθμός πολλαπλασιασμού παρατηρήθηκε σε έκφυτα σποροφύτων ηλικίας ενός έτους σε υπόστρωμα με 1 mg 1⁻¹ ΒΑ. Παρατηρήθηκε επίδραση της εποχής λήψης των εκφύτων στην αντίδρασή τους, καθώς έκφυτα που τοποθετήθηκαν στις αρχές Μαΐου παρουσίασαν υψηλότερο ρυθμό πολλαπλασιασμού σε σχέση με αυτά του Ιουλίου και Σεπτεμβρίου. Οι καλλιέργειες αναπτύχθηκαν σε άλατα του Woody Plant Medium (WPM), με βιταμίνες Mullin και 1.5 ή 3% σακχαρόζη. Διάφορες συγκεντρώσεις κυτοκινινών εξετάστηκαν για την επίδραση τους στον πολλαπλασιασμό των καλλιεργειών και ο υψηλότερος ρυθμός πολλαπλασιασμού επιτεύχθηκε σε υπόστρωμα με 1mg 1⁻¹ ΒΑ και 0.2 mg 1⁻¹ ΙΒΑ. Τα καλύτερα αποτελέσματα ριζοβολίας επιτεύχθηκαν μετά από καλλιέργεια σε 2 mg 1⁻¹ ΙΒΑ για μια εβδομάδα και στη συνέχεια σε υπόστρωμα χωρίς ορμόνες. Η εμβάπτιση της βάσης των μικροβλαστών σε πυκνά διαλύματα ΙΒΑ για διάφορες χρονικές περιόδους και στη συνέχεια καλλιέργεια τους in vitro σε WPM ή ex vitro σε έδαφος, δεν βελτίωσε την ριζοβολία τους, ενώ η χρήση της μεμβράνης sanitas ως κάλυψη των δοχείων καλλιέργειας είχε ευεργετικά αποτελέσματα στη ριζοβολία. Φυτάρια που τοποθετήθηκαν σε έδαφος από την Εύβοια εγκλιματίστηκαν σε υψηλότερο ποσοστό και είχαν καλύτερη ανάπτυξη συγκριτικά με αυτά που τοποθετήθηκαν σε μίγμα compost-περλίτη. Το αποτέλεσμα αυτό οφείλεται στην ύπαρξη μιας μυκόρριζας που βρέθηκε να υπάρχει στο έδαφος από την Εύβοια. Η μελέτη της επίδρασης των διάφορων υλικών κάλυψης έδειξε ότι αυτά επέδρασαν στη συγκέντρωση των αερίων και τη σχετική υγρασία εντός του δοχείου. Αυτό επηρέασε τόσο τα μορφολογικά όσο και τα ανατομικά χαρακτηριστικά των φύλλων που αναπτύχθηκαν. Στα δοχεία που καλύφθηκαν με ημίσκληρο πλαστικό, μεταλλικό ή σκληρό πλαστικό καπάκι παρατηρήθηκαν αλλαγές στη σύσταση των αέριων και υψηλή σχετική υγρασία, ενώ σχηματίστηκαν υπερενυδατομένα φύλλα με μεγάλα σφαιρικά στόματα που δύσκολα έκλειναν και προκαλούσαν γρήγορη αφυδάτωση των φύλλων και χαμηλή επιβίωση των φυταρίων κατά τον εγκλιματισμό. Υπό κάλυψη με μεμβράνη sanitas και magenta καπάκι, παρατηρήθηκε χαμηλή σχετική υγρασία και σχηματίστηκαν φύλλα με μικρά ελλειπτικά στομάτια, που έμοιαζαν με αυτά ex vitro αναπτυσσόμενων φυτών. Ομοίως ρίζες που σχηματίστηκαν υπό μεμβράνη sanitas είχαν ξύλωμα σε υψηλότερη αναλογία ως προς τη συνολική επιφάνεια της ρίζας από ρίζες που αναπτύχθηκαν υπό μεταλλικό καπάκι όπως αυτές ex vitro φυτών.


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).


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