Propagación in vitro de Psidium guajaba mediante organogénesis directa a partir de segmentos nodales

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 in vitro combinado con inducción rápida de brotes ex vitro para propagar árboles élite. La utilización de segmentos nodales permitió obtener en poco tiempo brotes adventicios adecuados para multiplicación masiva. La respuesta in vitro de los genotipos fue evaluada usando los medios de cultivo MS (Murashige y Skoog, 1962), Mc (Mascarenhas) y WPM (Woody Plant Medium) suplementados con 0,1 mg•L-1 de ácido indolácetico (AIA) y 0,25 mg•L-1 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-1 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 in vitro 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. In vitro propagation of Psidium guajaba using direct organogenesis from nodal segmentsMultiple 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.

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

Plants ◽

10.3390/plants9121657 ◽

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.

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588 PB 129 IN VITRO MULTIPLICATION OF STEWARTIA MALACODENDRON L., AN ENDANGERED WOODY SPECIES

HortScience ◽

10.21273/hortsci.29.5.516b ◽

1994 ◽

Vol 29 (5) ◽

pp. 516b-516

Author(s):

Murdock Ray Gillis ◽

Michael E. Kane

Keyword(s):

Woody Plant ◽

Unit Cost ◽

Woody Species ◽

Shoot Multiplication ◽

Shoot Tips ◽

Nodal Segments ◽

Deciduous Tree ◽

Landscape Plant ◽

Woody Plant Medium

Stewartia malacodendron L. (silky camellia), a small deciduous tree bearing showy flowers, has potential as a landscape plant. Propagation problems, limited availability and consequent high unit cost have slowed its acceptance as a landscape plant. Given its potential value as a landscape plant, studies were initiated to define a micropropagation protocol. Surface sterilized shoot tips and nodal explants from two-year-old container grown seedlings were established on Woody Plant Medium supplemented with 4.44 μM benzyladenine (BA) and solidified with 0.8% TC® Agar. Sustained growth of subcultured shoot tips and nodal segments required the addition of 8-15 μM gibberellic acid (GA3) to the medium. Regenerated shoots were 3 - 5 cm long, unbranched and typically consisted of three subdivisions. Effects of cytokinin type (BA, 2iP or kinetin) and concentration (0-25 μM) with factorial combinations of GA3 (0-30 μM) on shoot multiplication, elongation and diameter were determined after a 28 day culture period. Moderate GA, levels (10 & 20 μM) in combination with 2.5 μM BA yielded the highest quality microcuttings.

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In vitro propagation of Xao tam phan (Paramignya trimera (Oliv.) Guill.)

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v1it2.452 ◽

2017 ◽

Vol 1 (T2) ◽

pp. 48-57

Author(s):

Hieu Trung Tran ◽

Chung Van Huynh ◽

Hue Thi Linh Bui ◽

Ngan Thi My Luong ◽

Anh Lan Bui ◽

...

Keyword(s):

In Vitro Propagation ◽

Word Of Mouth ◽

Woody Plant ◽

Leaf Explants ◽

Anticancer Properties ◽

Best Response ◽

Woody Plant Medium ◽

Old Trees ◽

Cut Surface

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.

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In Vitro Propagation of Oriental White Oak Quercus aliena Blume

Forests ◽

10.3390/f10060463 ◽

2019 ◽

Vol 10 (6) ◽

pp. 463

Author(s):

Qiansheng Li ◽

Mengmeng Gu ◽

Min Deng

Keyword(s):

Woody Plant ◽

Germplasm Conservation ◽

Shoot Tips ◽

Nodal Segments ◽

Adventitious Bud ◽

White Oak ◽

Widespread Species ◽

Aseptic Culture ◽

Woody Plant Medium

Quercus aliena Blume, also known as the oriental white oak, is a widespread species in temperate forests of East Asia with significant ecological and economical importance. Establishing an efficient vegetative propagation system is important for its germplasm conservation and breeding program. Protocols of micropropagation from shoot tips and nodal segments were investigated in order to produce uniform high-quality seedlings. Nodal segments from 18 month old seedlings were used as explants to initiate the aseptic culture. The highest bud proliferation was achieved by subculturing the explants on 1/2 strength woody plant medium (WPM) with 2.0 mg·L−1 BA. WPM with 0.5 mg·L−1 BA and 0.05 mg·L−1 IBA was the best medium for subculture to obtain the vigorous regenerated shoots in this experiment. Nodal segments without shoot tips had a higher adventitious bud proliferation rate than those with shoot tips. The highest rate (41.5%) of rooting in vitro was induced by using WPM with 1.0 mg·L−1 IBA and 5 g·L−1 activated charcoal. Ex vitro rooting by dipping the proliferated shoots with 500 mg·L−1 IBA solution, then transplanting directly to potting mix with 50% peat and 50% horticultural perlite fostered the highest rooting percentage and survival rate of the plantlets.

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In vitro propagation of Epacris impressa (Epacridaceae) and the effects of clonal material

Australian Journal of Botany ◽

10.1071/bt98026 ◽

2000 ◽

Vol 48 (2) ◽

pp. 215 ◽

Cited By ~ 3

Author(s):

J. Anthony ◽

C. B. McLean ◽

A. C. Lawrie

Keyword(s):

In Vitro Propagation ◽

Root Formation ◽

Woody Plant ◽

Multiple Shooting ◽

Naphthaleneacetic Acid ◽

Root Induction ◽

Main Determinant ◽

Woody Plant Medium ◽

Half Strength

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.

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

Canadian Journal of Forest Research ◽

10.1139/x93-231 ◽

1993 ◽

Vol 23 (9) ◽

pp. 1821-1825 ◽

Cited By ~ 20

Author(s):

Snorri Baldursson ◽

Peter Krogstrup ◽

Jens Viktor Nørgaard ◽

Sven Bode Andersen

Keyword(s):

Chromosome Number ◽

Growth Regulator ◽

Woody Plant ◽

Adventitious Shoots ◽

Microspore Embryogenesis ◽

Naphthaleneacetic Acid ◽

Chromosome Counts ◽

Low Frequencies ◽

Woody Plant Medium

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.

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Micropropagation of Antelope Bitterbrush [Purshia tridentata (Pursh) DC.]

HortScience ◽

10.21273/hortsci.32.2.312 ◽

1997 ◽

Vol 32 (2) ◽

pp. 312-314 ◽

Cited By ~ 1

Author(s):

John L. Edson ◽

David L. Wenny ◽

Annette Leege-Brusven

Keyword(s):

Woody Plant ◽

Naphthaleneacetic Acid ◽

Ex Vitro ◽

Indolebutyric Acid ◽

Axillary Shoots ◽

Full Strength ◽

Woody Plant Medium ◽

Half Strength ◽

In Vitro Survival

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).

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Micropropagation of Chinquapin (Castanea henryi) Using Axillary Shoots and Cotyledonary Nodes

HortScience ◽

10.21273/hortsci13292-18 ◽

2018 ◽

Vol 53 (10) ◽

pp. 1482-1486 ◽

Cited By ~ 2

Author(s):

Huan Xiong ◽

He Sun ◽

Feng Zou ◽

Xiaoming Fan ◽

Genhua Niu ◽

...

Keyword(s):

Adventitious Shoots ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Nodal Segments ◽

Direct Organogenesis ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Axillary Shoots ◽

Cotyledonary Nodes

Castanea henryi is an important woody grain tree species native to China. The objective of the current study was to find the suitable plant growth regulators (PGRs) and the optimal concentrations for direct organogenesis by using axillary shoots and cotyledonary nodes. Seeds were collected from the field, sterilized, and germinated in vitro. Axillary shoots and cotyledonary nodes of 3-week-old seedlings were used as explants. To find the suitable PGR for adventitious shoot induction, 0.5 mg·L–1 6-benzylaminopurine (6-BA), 0.1 mg·L–1 indole-3-acetic acid (IAA), 0.1 mg·L–1 2,4-dichlorophenoxyacetic acid (2,4-D), or 0.1 mg·L–1 1-naphthaleneacetic acid (NAA) was supplemented to Murashige and Skoog (MS) medium containing 0.65% agar and 3% sucrose. A high induction percentage of adventitious shoots (85.67%) was obtained from cotyledonary nodes supplemented with 0.1 mg·L–1 2,4-D. The type of explant influenced shoot proliferation rates and quality. Apical explants produced more and longer shoots than nodal segments. For shoot multiplication, 1 mg·L–1 6-BA + 0.05 mg·L–1 indole-3-butyric acid (IBA) supplemented with MS medium produced 12.33 and 6.25 shoots per explant, respectively, from apical and nodal explants. For shoot elongation and strengthening, 2 mg·L–1 6-BA + 0.05 mg·L–1 IBA supplemented with MS medium was the best combination, producing shoots with a mean length of 3.50 cm, a diameter of 0.46 cm, and about eight leaves per shoot. The greatest rooting of 76.70% and 11.33 roots per shoot was achieved when cultured in MS medium supplemented with 3.5% perlite + 1.5 mg·L–1 IBA. For acclimatization of the rooted plantlets in the greenhouse, a survival rate of 80% was achieved. This protocol—from multiplication to acclimation—is helpful to realize mass propagation of high-quality trees of chinquapin for increasing production and nut quality.

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Διερεύνηση της επίδρασης της φυσιολογίας του εκφύτου, του υποστρώματος και των συνθηκών περιβάλλοντος στην οργανογένεση και τον εγκλιματισμό του quercus euboica

10.12681/eadd/18664 ◽

2008 ◽

Author(s):

Επαμεινώνδας Κάρτσωνας

Keyword(s):

Woody Plant ◽

Ex Vitro ◽

Woody Plant Medium

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.

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