In vitro regeneration of Pinus pinaster adult trees

2008 ◽  
Vol 38 (10) ◽  
pp. 2607-2615 ◽  
Author(s):  
N. De Diego ◽  
I. A. Montalbán ◽  
E. Fernandez de Larrinoa ◽  
P. Moncaleán
Keyword(s):  
Pinus Pinaster ◽  
In Vitro Regeneration ◽  
Forest Tree ◽  
Axillary Shoots ◽  
Breeding Programs ◽  
Practical Applications ◽  
Adult Trees ◽  
Improvement Programs ◽  
Conifer Trees

Regeneration of adult conifer trees by means of in vitro culture has been the subject of intense study during the last 20 years. Propagation by tissue culture may become the best method for obtaining multiple identical trees and for capturing the genetic gain in breeding programs. However, the method has several problems related to phase change of trees (juvenile–adult) that limit its practical applications. In this study, shoot buds were collected from 20 maritime pine ( Pinus pinaster Ait.) adult trees (>20 years old) from November to March. Buds were cut transversely into 0.5–1.0 cm slices and cultured on several media (DCR, WP and modified LP) supplemented with cytokinins (6-benzyladenine, metatopolin, or zeatin) at two concentrations (25 or 50 μmol/L). The highest organogenic response (axillary shoots formation ability) occurred on DCR medium supplemented with 25 μmol/L zeatin and metatopolin, and 25 or 50 μmol/L 6-benzyladenine. All shoots that regenerated on DCR medium with 25 μmol/L 6-benzyladenine developed healthy and well rooted plantlets. The ability to micropropagate adult trees represents a significant progress in the application of biotechnology to forest tree improvement programs, and it opens the possibility of using select trees in agroforestry areas under biotic or abiotic stress.

scholarly journals Date palm micropropagation: Advances and applications

2017 ◽  
Vol 41 (4) ◽  
pp. 347-358 ◽  
Author(s):  
Jameel Mohammed Al-Khayri ◽  
Poornananda Madhava Naik
Keyword(s):  
Somaclonal Variation ◽  
Large Scale ◽  
Date Palm ◽  
In Vitro Regeneration ◽  
Phoenix Dactylifera ◽  
Climatic Conditions ◽  
Chemical Components ◽  
Palm Trees ◽  
Adult Trees

ABSTRACT Date palm (Phoenix dactylifera L.) is a fruit tree resilient to adverse climatic conditions predominating in hot arid regions of the Middle East and North Africa. The date fruit contains numerous chemical components that possess high nutritional and medicinal values. Traditional propagation by offshoots is inefficient to satisfy current demands for date palm trees. Alternatively, micropropagation provides an efficient means for large-scale propagation of date palm cultivars. Both somatic embryogenesis and organogenesis, either directly or indirectly though the callus phase, have been demonstrated in date palm in vitro regeneration. Culture initiation commonly utilizes shoot-tip explants isolated from young offshoots. Recently, the immature inflorescences of adult trees were utilized as an alternative nondestructive source of explants. In addition to the nature of the explant used, successful plant regeneration depends on the cultivar, composition of the culture medium and physical status. Challenges of date palm micropropagation include long in vitro cycle, latent contamination, browning, somaclonal variation as well as ex vitro acclimatization and transplanting. A remarkable amount of research investigating these factors has led to optimized protocols for the micropropagation of numerous commercially important cultivars. This has encouraged the development of several international commercial tissue culture laboratories. Molecular characterization provides an assurance of genetic conformity of regenerated plantlets, a key feature for commercial production. This article describes date palm micropropagation protocols and also discusses recent achievements with respect to somaclonal variation, molecular markers, cryopreservation and future prospects.

scholarly journals A Novel Method for Efficient Micropropagation of Radermachera xylocarpa (Roxb.) K. Schum., a Rare Medicinally Important Forest Species

2013 ◽  
Vol 23 (1) ◽  
Author(s):  
Manjary Sathe ◽  
Megha Vibhute ◽  
Monica Jain ◽  
Pankaj Srivastav
Keyword(s):  
Tree Species ◽  
Large Scale ◽  
In Vitro Regeneration ◽  
Forest Tree ◽  
Root Induction ◽  
Mass Propagation ◽  
Forest Tree Species ◽  
Bud Induction ◽  
Indigenous Forest

Radermachera xylocarpa (Roxb.) K. Schum. is a rare indigenous forest tree species which is utilized for its wood and medicinal properties. Due to its overexploitation and specific habitat requirements the species is restricted to limited areas. In vitro mass propagation of tree species faces various challenges and no such efforts have yet been taken in propagation of this useful plant using these methods.  In order to overcome the hurdles and understanding an urgent need of its conservation and mass propagation present authors attempt to develop a simple effective tissue culture protocol for regeneration of R. xylocarpa. Nodal explants were cultured on MS supplemented with various concentrations of cytokinins and auxins.  Among different cytokinins, maximum bud induction and proliferation was obtained in media supplemented with Kn along with IBA and for effective root induction which is tough to obtain in tree species, 100% rooting was achieved in cultures with increasing concentrations of IBA. Field survival is a major challenge with regenerated plants of forest tree species. We report here for the first time 100% survival of plants in soil by carefully standardizing the period of hardening and acclimatization procedures. A novel and effective in vitro regeneration protocol of R. xylocarpa has been successfully standardized which can be adopted for large scale propagation, reforestation and conservation of rare Radermachera xylocarpa of medicinal importance.Plant Tissue Cult. & Biotech. 23(1): 21?29, 2013 (June)DOI: http://dx.doi.org/10.3329/ptcb.v23i1.15556

scholarly journals Effect of Plant Growth Regulators (BA, KIN and NAA) on In vitro Propagation of Papaya (Carica papaya)

2020 ◽  
pp. 15-23
Author(s):  
Nahida Hasan ◽  
Humayra Huq ◽  
Fahima Khatun ◽  
Shamim Ara Sumi
Keyword(s):  
In Vitro Propagation ◽  
Carica Papaya ◽  
In Vitro Regeneration ◽  
Combined Treatment ◽  
Survival Rates ◽  
Tween 20 ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Breeding Programs

The present research was carried out in Biotechnology Laboratory of the Department of Biotechnology, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar, Dhaka-1207 from the period of September 2017 to June 2018. This research aims to study the effect of Benzyladenine (BA), Kinetin (KIN) and Naphthalene acetic acid (NAA) either in combination or alone on In vitro propagation of papaya (Carica papaya). The shoot tips of young shoots were used as explant, which was sterilized using freshly prepared 0.1% HgCl2 mixing with few drops of Tween-20, were inoculated in MS media supplemented with 0.1% activated charcoal. The minimum days to shoot induction (10.25) were recorded on MS medium containing 0.5 mg/L BA. The highest shoots (4.5) and length of shoot (5.75 cm) observed in 1.0 mg/L BA. The combined treatment 1.0 mg/L BA+0.75 mg/LKIN gave the highest number of shoots (5.25) and length of shoot (5.78 cm).The minimum days (8.5) to root induction was reported in 2.0 mg/L NAA along with maximum 8.25 roots per plantlet. The highest length of root (6.92 cm) was observed in 2.0 mg/L NAA. In regenerated plantlets, 80% survival rates were observed in growth chamber conditions and 75% in the open atmosphere were achieved. Finally, the in vitro regeneration protocol described herein can potentially be used as a tool in molecular breeding programs for the improvement of different cultivars and genotypes of papaya.

scholarly journals Micropropagation of Salvia broussonetii Benth. - A Medicinal Plant Species

1970 ◽  
Vol 16 (1) ◽  
pp. 19-23 ◽  
Author(s):  
S Mederos-Molina
Keyword(s):  
Ascorbic Acid ◽  
Medicinal Plant ◽  
Plant Species ◽  
Culture Medium ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Axillary Shoots ◽  
Half Strength ◽  
Important Medicinal Plant

The purpose of this study was to establish culture medium requirements for micropropagation of Salvia broussonetii Benth., an important medicinal plant. Cultures were initiated from axillary shoots collected from mature plants. Most satisfactory results were achieved using a MS.2 medium supplemented with 1 mM ascorbic acid, 1.44 µM GA3 and 1.11 µM BAP. Axillary nodes were used for in vitro regeneration of multiple shoots and best results were achieved with MS.2 medium plus 1.44 µM GA3, 2.66 µM BAP and 1.14 µM IAA. Shoots rooted without symptoms of chlorosis or necrosis in half-strength MS.2 medium plus 1.44 µM GA3 and 2.28 µM IAA.Key words: Axillary shoots, Micropropagation, Medicinal plant, Salvia broussonetiiDOI = 10.3329/ptcb.v16i1.1101Plant Tissue Cult. & Biotech. 16(1): 19-23, 2006 (June)

scholarly journals Response of root explants to in vitro cultivation of marketable garlic cultivars

2013 ◽  
Vol 31 (1) ◽  
pp. 80-85 ◽  
Author(s):  
Danielle C Scotton ◽  
Vagner Augusto Benedito ◽  
Jeanne B de Molfetta ◽  
Benedita Inês FP Rodrigues ◽  
Augusto Tulmann-Neto ◽  
...  
Keyword(s):  
Disease Transmission ◽  
In Vitro Regeneration ◽  
Growth Conditions ◽  
Production Costs ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Root Explants ◽  
Breeding Programs

Garlic cultivars are sexually sterile under standard growth conditions, with direct implications for commercial production costs as well as breeding programs. Garlic is propagated commercially via bulblets, which facilitates disease transmission and virus load accumulation over vegetative generations. Tissue culture produces virus-free clones that are more productive, while keeping the desired traits of the cultivar. Consequently, this technique allows studies of garlic genetics as well as guarantees genetic conservation of varieties. We aimed at analyzing the in vitro regeneration of eight marketable cultivars of garlic using root segments as explants. For each genotype, bulblet-derived explants were isolated and introduced into MS medium supplemented with 2,4-D and 2-iP. Calli were transferred to MS medium supplemented with 8.8 mM BAP and 0.1 mM NAA (regeneration medium A), or with 4.6 mM kinetin alone (regeneration medium B). The calli were then evaluated for regeneration frequency after sixty days of in vitro cultivation. The noble cultivar 'Jonas' presented the highest rates of plant regeneration among the cultivars tested. The medium A, which contained auxin and cytokinin, induced the highest regeneration rates of all cultivars. The process described herein is simple, reproducible and can potentially be used as a tool in molecular breeding strategies for other marketable cultivars and genotypes of garlic.

scholarly journals Regeneration of Pinus halepensis (Mill.) through Organogenesis from Apical Shoot Buds

Forests ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 363
Author(s):  
Cátia Pereira ◽  
Itziar A. Montalbán ◽  
Ana Pedrosa ◽  
Jéssica Tavares ◽  
Alexey Pestryakov ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Pinus Halepensis ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Embryogenic Calli ◽  
Shoot Buds ◽  
Apical Shoot ◽  
Adult Trees

Organogenesis and somatic embryogenesis have been widely applied as the two main regeneration pathways in plant tissue cultures. However, recalcitrance is still the main restriction in the clonal propagation of many woody species, especially in conifers. They undergo a “phase change” that leads to significant loss of vegetative propagation capacity, reducing the aptitude of tissues and organs to be regenerated in vitro beyond this point. In line with this, the in vitro regeneration of mature conifer trees has been a long-cherished goal in many laboratories worldwide. Based on previous works in Pinus species regeneration from adult trees, we now present data about the culture of apical shoot buds in an attempt to induce organogenesis and somatic embryogenesis to clone mature trees of Aleppo pine (Pinus halepensis). Reinvigorated axillary shoots were submitted to conditions usually applied to induce somatic embryogenesis through the manipulation of culture media, including the use of auxins such as 2,4-Dichlorophenoxyacetic acid and 1-Naphthaleneacetic acid, cytokinins (6-benzyladenine and kinetin), and phytosulfokine (50, 100, and 200 nM). Although somatic embryos could not be obtained, an embryogenic-like tissue was produced, followed by the emergence of actively proliferating non-embryogenic calli. Variations in the consistence, texture, and color of non-embryogenic calli were observed; especially those arising in the media containing phytosulfokine. Reinvigorated shoots, induced by 22 or 44 µM 6-benzyladenine, were obtained through organogenesis and acclimatized, and phenotypically normal plants were obtained.

In vitro regeneration from leaf-base segments in three genotypes of Urochloa spp.

2018 ◽  
Vol 69 (5) ◽  
pp. 527 ◽  
Author(s):  
Diliane Harumi Yaguinuma ◽  
Luciana Midori Takamori ◽  
Adriana Mendonça de Oliveira ◽  
Luiz Gonzaga Esteves Vieira ◽  
Alessandra Ferreira Ribas
Keyword(s):  
In Vitro Regeneration ◽  
Forage Grasses ◽  
Leaf Base ◽  
Breeding Programs ◽  
Seed Formation ◽  
Regenerated Plants ◽  
Tropical Forage ◽  
Half Strength ◽  
The Media

The key agricultural species of Urochloa P.Beauv. (signal grass), important as tropical forage grasses, are characterised by asexual seed formation (apomixis), and this presents a challenge for breeding programs. Biotechnological approaches could be an option to develop improved cultivars. We evaluated the regenerative potential from three commercial genotypes, U. brizantha cv. Marandu, U. decumbens cv. Basilisk and U. ruziziensis cv. Ruziziensis, by using leaf-base segments as explants. We tested two auxins (2,4-D and picloram) and one cytokinin (TDZ) at four concentrations (1, 2, 3 and 4 mg L–1). Seeds were scarified, peeled and disinfected before inoculation on half-strength MS media in the dark for 14 days. Leaf-base explants were sectioned in thin slices and inoculated into the media. We analysed the number of primary calluses, number of calluses with shoots clusters and the average of regenerated plants. The lowest concentration of auxins tested (1 mg L–1) yielded the highest number of regenerated plants for Marandú and Basilisk, whereas the optimum for Ruziziensis was 2 mg L–1. Medium with higher concentrations of TDZ (4 mg L–1) was required to produce high frequency of plants for all genotypes. Explants cultured on media with TDZ produced very few calluses. These results indicate that the auxins and cytokinin tested can induce plant regeneration from Urochloa leaf-base segments, and may be used to produce transgenic plants in genetic transformation studies.

scholarly journals In Vitro Techniques to the Conservation and Plant Regeneration of Malanga (Colocasia esculenta L. Schott)

HortScience ◽  
2019 ◽  
Vol 54 (3) ◽  
pp. 514-518 ◽  
Author(s):  
Eucario Mancilla-Álvarez ◽  
Marco A. Ramírez-Mosqueda ◽  
Samantha Arano-Avalos ◽  
Rosalía Núñez-Pastrana ◽  
Jericó J. Bello-Bello
Keyword(s):  
Genetic Resource ◽  
In Vitro Regeneration ◽  
Colocasia Esculenta ◽  
In Vitro Conservation ◽  
Plant Genetic Resource ◽  
Minimal Growth ◽  
Shoot Number ◽  
Number Of Leaves ◽  
Improvement Programs

Malanga (Colocasia esculenta) is a plant genetic resource that requires biotechnological strategies for conservation and propagation. One time-, labor-, and space-saving option is in vitro conservation and regeneration. The objective of this study was to develop a protocol for in vitro regeneration and conservation of germplasm of C. esculenta var. criolla. For conservation through minimal growth, we assessed several concentrations of Murashige and Skoog (MS) medium (one-third, one-half, and three-quarter strength), the growth retardant ancymidol (0, 1, 2, and 3 mg·L−1), and the osmoregulator polyethylene glycol (PEG-8000 mw) at different concentrations (0, 10, 20, and 30 g·L−1). For in vitro conservation, the percent survival, shoot number and length, and number of leaves and roots per explant were evaluated after 24 weeks. For in vitro regeneration, different concentrations of thidiazuron (TDZ: 0, 0.5, 1, 1.5, and 2 mg·L−1) and 6-benzylaminopurine (BAP; 0, 1, 2, 3, and 4 mg·L−1) were evaluated. After 4 weeks of cultivation, the percent response, shoot number, and number of leaves per explant were recorded. During in vitro conservation, it was noted that the treatment including 2 mg·L−1 ancymidol resulted in a retarded development, without affecting the survival of the C. esculenta germplasm. With regard to shoot regeneration, 7.60 shoots per explant were obtained using 2 mg·L−1 TDZ. Finally, 98% survival was achieved during the acclimatization process. This study will contribute to the establishment of genetic improvement programs through in vitro conservation and propagation of this valuable plant genetic resource.

scholarly journals The effect of auxins in inducing organogenesis or somatic embryogenesis in mature sunflower zygotic embryo derived apex

2020 ◽  
Vol 48 (1) ◽  
pp. 150-161
Author(s):  
Adriana AURORI ◽  
Imola MOLNAR ◽  
Elena RAKOSY-TICAN
Keyword(s):  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Mature Stage ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Axillary Shoots ◽  
Mature Embryos ◽  
Cytokinin Treatment ◽  
2,4 Dichlorophenoxyacetic Acid

Induction of shoots or of somatic embryos is the key step for gaining the morphogenetic potential in sunflower (Helianthus annuus L.), species known as recalcitrant to in vitro regeneration. In the immature zygotic embryo derived tissues or in other juvenile tissues resulted from seedlings, the acquisition of the competence for regeneration can be achieved directly by cytokinin treatment or by preconditioning the explants on cytokinin containing medium. In this paper is presented a new type of explant for sunflower in vitro culture, consisting of the apex with primordial leaves, resulted from ungerminated mature zygotic embryo, in which a specific morphogenetic response was triggered by the exogenously applied auxins. Among the auxins tested, indole-3-acetic acid, indole-3-butyric acid and 1-naphthaleneacetic acid are inducers of an organogenetic response, apical/axillary shoots and adventitious buds being regenerated while 2,4-dichlorophenoxyacetic acid, 3,6-dichloro-2-methoxybenzoic acid and 4-amino-3,5,6-trichloropicolinic acid led to somatic embryo formation. Among the auxins tested only 4-amino-3,5,6-trichloropicolinic acid sustains the embryos development up to mature stage. A high amount of sucrose (120 g L-1) supplied during the auxin treatment promotes the maturation of the embryos directly on the induction medium for all tested auxins with embryogenic effect. These findings show that regardless of the type of morphogenetic response aimed in sunflower meristematic tissues resulted from mature embryos, the presence of auxins is mandatory.

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