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

2021 ◽  
Author(s):  
Cátia Pereira ◽  
Itziar A. Montalbán ◽  
Ana Pedrosa ◽  
Jéssica Tavares ◽  
Alexey Pestryakov ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Pinus Halepensis ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Embryogenic Calli ◽  
Mature Trees ◽  
Axillary Shoots ◽  
Shoot Buds ◽  
Apical Shoot ◽  
Adult Trees

Organogenesis and somatic embryogenesis have been widely applied as the two main regeneration pathways in plant tissue culture. However, recalcitrance is still a main restriction in the clonal propagation of many woody species, especially in conifers. They undergo a “phase change” that leads to significant loss of organogenic and embryogenic capacity, thus reducing the responsive tissues or organs to juvenile material, and narrowing the competence window. In this sense, in vitro regeneration of mature conifer trees has been a long-cherished goal in many laboratories worldwide. In this work, apical shoot buds were used as explants for both organogenesis and somatic embryogenesis in order to cloning mature trees of Aleppo pine. Reinvigorated axillary shoots were submitted to somatic embryogenesis induction through the manipulation of culture media, including the use of auxins such as 2,4-D and NAA, cytokinins (BA and kinetin) and phytosulfokine (50, 100 and 200 nM). Although somatic embryos could not be obtained, embryogenic-like tissue was produced followed by the appearance of actively proliferating non-embryogenic calli and differences between treatments were found, especially when phytosulfokine was added to the induction media. Organogenic system produced reinvigorated shoots from both BA treatments tested (22 and 44 µM), from juvenile somatic trees and adult trees, and ex-vitro acclimatized plants were developed.

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.

Evaluation of the Enhanced Regeneration System for in vitro regeneration in barley

2006 ◽  
Vol 86 (1) ◽  
pp. 63-69
Author(s):  
Seedhabadee Ganeshan ◽  
Brian J Weir ◽  
Monica Båga ◽  
Brian G Rossnagel ◽  
Ravindra N Chibbar
Keyword(s):  
Hordeum Vulgare ◽  
Embryogenic Callus ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Cold Treatment ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Hordeum Vulgare L ◽  
Best Response

A simple two-step model for evaluation of in vitro regeneration protocols is proposed based on callus induction and regeneration from immature scutella of two Canadian barley (Hordeum vulgare L.) genotypes, AC Metcalfe and SB92559 using the Enhanced Regeneration System (ERS). The number of explants producing embryogenic callus, the number of plants per embryogenic callus and the number of plants per explant were considered. Tissue culture parameters included three combinations of growth regulators, two carbon sources in culture media, and three cold treatment regimes of spikes prior to scutella isolation. Culture medium containing 5 µM 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 µM benzyl adenine (BA) induced the highest percent of embryogenic calli and the highest number of shoots per embryogenic callus from AC Metcalfe. Medium containing 3.75 µM 2,4-D and 0.75 µM BA gave the best response for SB92559. Both genotypes produced more shoots on maltose than on sucrose medium. A 2-d treatment of spikes at 4°C resulted in best response for SB92559. Regeneration response from AC Metcalfe scutella from spikes was unaffected by being subjected to 2, 4 or 6 d of cold. Conditions resulting in best responses from both genotypes were tested on four commercial barley varieties. However, these lines showed inferior regeneration compared to SB92559 and AC Metcalfe. Key words: Hordeum vulgare, scutella, embryogenic callus, shoot production

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 Effect of culture media, auxins and genotypes on plantlet regeneration from oil palm (Elaeis guineensis Jacq.) zygotic embryos through somatic embryogenesis

2021 ◽  
Vol 42 (5) ◽  
pp. 1232-1238
Author(s):  
D.S. Sparjanbabu ◽  
◽  
P.N. Kumar ◽  
S.R.K. Motukuri ◽  
D. Ramajayam ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Oil Palm ◽  
Callus Induction ◽  
Elaeis Guineensis ◽  
Culture Media ◽  
Plantlet Regeneration ◽  
Zygotic Embryos ◽  
Embryogenic Calli ◽  
Significant Difference

Aim: This study evaluated efficient culture media for the regeneration of elite material through somatic embryogenesis from oil palm zygotic embryos. Methodology: For callus induction, zygotic embryos of four elite genotypes (G1-264T, G2-238DX17P, G3-37DX17P and G4-237T) were cultured on three basal media (Y3, MS and N6) with different auxin 2 mg l-1 (Picloram, 2,4-D and Dicamba) combinations. Subculture was made every month for three passages. It evaluated various callus characters. The embryogenic calli from callus induction media were transferred to the embryo maturation medium and subcultured until the polyembryoids formed. For shoot and root formation, somatic embryo clumps were transferred into regeneration media. In-vitro plantlets with well-grown roots were hardened in pots for six weeks and assessed clonal fidelity using polymorphic SSR primers. Results: Among the treatments, calli from N6+2,4-D, Y3+2,4-D and N6+Picloram showed the highest embryogenic callus potential. G4-237T induced more embryogenic calli (32.982) among genotypes, which was on par with G1-264T (24.196). Embryogenic calli grown on N6 media with Dicamba showed the highest proliferation rate (1.141). After 60 days of culture on regeneration media, the highest number of plantlets per somatic embryogenic clump was obtained from G1-264T on N6 media supplemented with Dicamba. Interpretation: Culture media salt concentration showed a significant difference among media by causing perturbations of auxin flow during somatic embryogenesis affecting callus induction, proliferation and plantlet regeneration. This may be useful for standardizing the genotype-specific regeneration media in oil palm.

scholarly journals The Role of Sequestrene 138 in Highbush Blueberry (Vaccinium corymbosum L.) Micropropagation

HortScience ◽  
2018 ◽  
Vol 53 (10) ◽  
pp. 1487-1493 ◽  
Author(s):  
Doina Clapa ◽  
Claudiu Bunea ◽  
Orsolya Borsai ◽  
Adela Pintea ◽  
Monica Hârța ◽  
...  
Keyword(s):  
Culture Media ◽  
Iron Concentration ◽  
Vaccinium Corymbosum ◽  
Carotenoid Content ◽  
Free Medium ◽  
Axillary Shoots ◽  
Different Types ◽  
The Media ◽  
Dark Green

The current research was carried out to investigate the effects of iron source in the culture media for Vaccinium corymbosum L. ʻBluerayʼ, ʻDukeʼ, and ʻPatriotʼ cultivars grown on five different types of medium (Woody Plant Medium supplemented with 1.0 mg·L−1 zeatin and 0, 25, 50, 75, and 100 mg·L−1 Sequestrene 138). After 10 weeks of culture, seven physiological parameters were measured, such as the number and length of axillary shoots, rooting and acclimatization percentage, as well as chlorophyll (a, b, a/b) and carotenoid content of the leaves. Adding Sequestrene 138 to the culture media led to a slight decrease of the proliferation rate but increased the length of the shoots. The chlorophyll and carotenoid content in all of the three cultivars was considerably increased as the iron concentration of the media increased. The shoots developed on the Sequestrene 138–free medium were chlorotic and short, whereas at different concentrations of iron in the culture medium the shoots were dark green and vigorous, providing a greater acclimatization success than those grown in iron-free medium.

scholarly journals Can Ethylene Inhibitors Enhance the Success of Olive Somatic Embryogenesis?

Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 168
Author(s):  
Muhammad Ajmal Bashir ◽  
Cristian Silvestri ◽  
Amelia Salimonti ◽  
Eddo Rugini ◽  
Valerio Cristofori ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Salicylic Acid ◽  
Silver Nitrate ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Cobalt Chloride ◽  
Zygotic Embryos ◽  
Ethylene Inhibitors ◽  
Embryogenic Calli ◽  
Stimulatory Action

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.

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