Prácticas del laboratorio de cultivo de células y tejidos vegetales

2021 ◽  
Author(s):  
Gloria Irma Ayala Astorga
Keyword(s):  
Woody Plant ◽  
Woody Plant Medium

Este manual está enfocado en la propagación asexual de los vegetales, por ende, las prácticas se orientan a la identificación de tipos de plantas, elaboración de medios de cultivo a partir de sales de Murashige y Skoog, como de Woody Plant Medium (WPM), así como la siembra de semillas in vitro y la organogénesis. Consta de un total de 13 prácticas acompañadas de figuras que ilustran la descripción del procedimiento, además de dos apéndices que guían la creación de medios de cultivo.

Period of harvest, sprouting ability of cuttings, and in vitro plant regeneration in Fraxinus excelsior

1994 ◽  
Vol 72 (2) ◽  
pp. 261-267 ◽  
Author(s):  
Conceição Eneida Silveira ◽  
Alain Cottignies
Keyword(s):  
Chromosome Number ◽  
In Vitro Culture ◽  
Adventitious Roots ◽  
Woody Plant ◽  
Fraxinus Excelsior ◽  
Stem Cuttings ◽  
Natural Conditions ◽  
Apical Bud ◽  
Woody Plant Medium

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.

scholarly journals In vitro establishment and proliferation of red currant cultivars

2012 ◽  
Vol 39 (No. 1) ◽  
pp. 21-25 ◽  
Author(s):  
J. Sedlák ◽  
F. Paprštein
Keyword(s):  
Woody Plant ◽  
Adventitious Bud ◽  
Mineral Salts ◽  
The Czech Republic ◽  
Shoot Number ◽  
Adventitious Bud Formation ◽  
Woody Plant Medium ◽  
In Vitro Establishment ◽  
Red Currant

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 Höllandische 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 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 Holländische that produced 2.1 ± 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.

Manipulation of desiccation-sensitive axes of wampee (Clausena lansium) to facilitate increased dehydration tolerance

2000 ◽  
Vol 10 (3) ◽  
pp. 397-400
Author(s):  
J.R. Fu ◽  
X.M. Huang ◽  
S.Q. Songa
Keyword(s):  
Butyric Acid ◽  
Water Loss ◽  
Adventitious Roots ◽  
Woody Plant ◽  
Dehydration Tolerance ◽  
Woody Plant Medium ◽  
Napthaleneacetic Acid ◽  
Pre Treatment

AbstractThe plumules of newly-excised wampee embryos, which are more sensitive to dehydration than the roots, became more resistant to water loss when axes were allowed to sprout on woody plant medium [WPM; McCown and Lloyd (1981) Hortscience16, 453] before being dried. Pre-treatment of sprouting axes (seedlings) with sucrose incorporated in the WPM enhanced survival. Although the roots withered following further dehydration of seedlings cultured on WPM containing 60% sucrose, excised plumules were capable of generating adventitious roots when a combination of 10 mM α-napthaleneacetic acid and 10 mM indole-3-butyric acid was used during subsequent in vitro incubation.

scholarly journals Micropropagation of sea buckthorn (Hippophae rhamnoides L.)

1995 ◽  
Vol 4 (5-6) ◽  
pp. 503-512
Author(s):  
Yingmou Yao
Keyword(s):  
Growth Regulator ◽  
Woody Plant ◽  
Average Rate ◽  
Root Nodules ◽  
Optimal Concentration ◽  
Sea Buckthorn ◽  
Adventitious Buds ◽  
Woody Plant Medium ◽  
Different Origins

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.

scholarly journals IN VITRO FLOWERING OF MINIATURE ROSE CULTIVARS

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 694e-694
Author(s):  
Michael Kane ◽  
Nancy L. Philman ◽  
Francis J. Marousky
Keyword(s):  
Woody Plant ◽  
Cut Flowers ◽  
Mineral Salts ◽  
Lateral Buds ◽  
Woody Plant Medium ◽  
Causative Agents ◽  
Ascorbic Acids ◽  
Indole 3 Acetic Acid ◽  
Stem Segments

Premature deterioration and/or wilting of cut flowers such as roses (“bent neck”) has been attributed to vascular blockage within the cut stem. Vascular blockage has been attributed to both the proliferation of bacteria in the cut flower water and/or to products exuded by the stem. Separation of these causative agents is prevented by the inability to obtain intact microbe-free flowers. With the objective to produce microbe-free flowers, 36 miniature rose cultivars were screened for their capacity to flower in vitro. Stem segments containing single lateral buds were surface sterilized in 1.05% (v/v) sodium hypochlorite and rinsed three times in sterile distilled deionized water. Buds were established on medium consisting of Murashige and Skoog mineral salts, Woody Plant Medium organics, 3.0% (w/v) sucrose, 0.5 mg/liter benzyladenine, 0.1 mg/liter indole-3-acetic acid, and 50 mg/liter each citric and ascorbic acids. Medium was solidified with 1.5 g/liter gelrite and 4 g/liter TC® agar. Of the 36 cultivars screened, eight (22%) grew poorly in vitro. Of the 28 responsive cultivars, 14 (50%) produced flower buds in vitro However, only six cultivars produced open flowers in vitro.

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

HortScience ◽  
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.

scholarly journals The in vitro shoot regeneration of Ehretia asperula Zol. and Mor.

2019 ◽  
Vol 2 (6) ◽  
pp. 75-83
Author(s):  
Le Thi Thuy Tien
Keyword(s):  
Woody Plant ◽  
Ms Medium ◽  
Benzyl Adenine ◽  
Shoot Initiation ◽  
In Vitro Shoots ◽  
Woody Plant Medium ◽  
Number Of Leaves ◽  
Growth Experiments ◽  
In Vitro Shoot Regeneration

Xa den young branches in the orchard were sterilized and used as explants for shoot initiation and growth experiments. The shoot induction was carried out with BA (benzyl adenine) or TDZ (thidiazuron). New shoots sprouted after one week of culture and the highest shoots (2.02 cm) were on MS medium (Murashige and Skoog medium) with BA 0.6 mg/L after 3 weeks. Furthermore, the number of leaves per shoot was also higher than other treatments (8.31 leaves per shoot). In vitro shoots were used in other experiments to investigate the effects of explants, biotin concentrations, minerals, type and concentration of cytokinins on the formation and elongation of shoots and clusters. When 5 mg/L biotin was added to MS medium, shoots grew better. The MS medium appeared to be most suitable for the initiation and elongating of shoots, followed by WPM (woody plant medium) and SH medium (Schenk and Hildebrandt medium), while B5 medium (Gamborg B5 medium) was the least effective. The spouting from three-week-old explants was earlier than others (4 and 5 weeks of age), which in turn affected on the shoot elongating. BA 1.5 mg/L was suitable to induce shoot clusters (3.91 shoots per explant) after 4 weeks.

scholarly journals PERBANYAKAN AKASIA HIBRIDA (Acacia mangium × Acacia auriculiformis) MELALUI SUBKULTUR BERULANG

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Yelnititis Yelnititis ◽  
Sri Sunarti
Keyword(s):  
Woody Plant ◽  
Basal Medium ◽  
Shoot Multiplication ◽  
Shoot Length ◽  
Acacia Auriculiformis ◽  
Shoot Induction ◽  
Benzyl Adenine ◽  
Single Node ◽  
Woody Plant Medium

In vitro culture is a promising technique for mass propagation of high-value species. Study of propagation for Acacia hybrid (A. mangium x A. auriculiformis) through this technique has been conducted using single node stem from seedlings as explants. Growth medium used was modified Murashige and Skoog (MS), basal medium Woody Plant Medium (WPM), and Gamborg (B5) supplemented. The study was conducted in two stages, namely shoot induction and shoot multiplication. The treatment tested was the Benzyl Adenine (BA) supplementation at the concentration of 0.3; 0.7; and 1.0 mgL-1 of. Observation was conducted on the frequency of shoot induction, number of shoot, shoot length and visual performance of the culture. The result showed that treatment of BA 0.7 mgL-1 on modified MS medium is the best for shoot induction, shoot multiplication and visual performace of the culture. The average of number of shoot was 2.6; 5.0 and 7.7 shoots on the first three consecutive subcultures. Changing to different basal medium on the fourth subculture showed that the treatment of BA 0.7 mgl-1 is the best condition for shoot regeneration (12.60 shoots) and shoot length (6.97 cm). The culture from this treatment showed the best visual morphological performance.Keywords:Acacia hybrid; multiplication; subculture; in vitro; BA. ABSTRAKKultur in vitro merupakan suatu teknik yang menjanjikan untuk perbanyakan massal spesies-tanaman bernilai tinggi. Penelitian perbanyakan akasia hibrid (A. mangium x A. auriculiformis) melalui kulturin vitro telah dilakukan dengan menggunakan eksplan berupa batang satu buku yang berasal dari anakan. Media tumbuh yang digunakan adalah media dasar Murashige dan Skoog (MS) yang sudah dimodifikasi, media dasar Woody Plant Medium (WPM), dan Gamborg (B5). Penelitian dilakukan dalam dua tahap yaitu induksi tunas dan perbanyakan tunas. Perlakuan yang diuji adalah penggunaan Benzyl Adenine (BA) dengan konsentrasi 0,3; 0,7 dan 1,0 mg L-1. Pengamatan dilakukan terhadap waktu induksi tunas, jumlah tunas, tinggi tunas dan penampilan biakan secara visual. Hasil penelitian menunjukkan bahwa penggunaan BA 0,7 mg L-1 pada media MS modifikasi merupakan perlakuan terbaik untuk induksi tunas, perbanyakan tunas, tinggi tunas, dan kondisi biakan secara visual. Jumlah rata-rata tunas yang dihasilkan dari perlakuan ini adalah 2,6; 5,0 dan 7,7 tunas pada subkultur pertama, kedua dan ketiga. Pada penggunaan media dasar berbeda pada subkultur keempat menunjukkan bahwa perlakuan BA 0,7 mg L-1 merupakan perlakuan terbaik dengan jumlah tunas sebanyak 12,60 tunas dan rata-rata tinggi tunas 6,97 cm. Biakan yang dihasilkan dari perlakuan tersebut mempunyai penampilan yang baik dan normal.

scholarly journals 100 A Comparison of Two Media on Growth of Deciduous Azalea Shoots In Vitro

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 458E-458
Author(s):  
Carol D. Robacker ◽  
Betty Robicheaux
Keyword(s):  
Woody Plant ◽  
Culture Media ◽  
Shoot Tips ◽  
Fresh Medium ◽  
First Flush ◽  
Wide Range ◽  
Woody Plant Medium ◽  
Made In

Micropropagation is a useful technique to propagate species such as deciduous azaleas, which are difficult to root from cuttings. To develop a micropropagation protocol that would be effective with a wide range of species and cultivars of native azalea, two culture media, Woody Plant Medium (WPM) (Lloyd and McCown, 1980) and ER medium (Economou and Read, 1994) were evaluated for ability to support growth of 11 species and four cultivars of deciduous azalea. Shoot tips were obtained from the first flush of growth in the spring on plants growing in the greenhouse or field. Following disinfection, the terminal and basal ends were removed from each explant. The explants were placed in culture tubes containing either WPM or ER medium with 12 mg/L 2iP and solidified with agar. Cultures were transferred to fresh medium every 4 to 6 weeks. Initial evaluations were made in 1996, and the experiment was repeated in 1997. In 1998, six of the taxa were evaluated for a third year. For most of the taxa evaluated, growth was superior on ER medium. On WPM, many of the cultures browned and died. R. canescens, R. viscosum, R. prunifolium, and R. austrinum are examples of species that preferred ER medium. R. alabamense, R. arborescens, and `My Mary' performed similarly on either medium.

scholarly journals Plantlets from encapsulated shoot buds of Catalpa ovata G. Don

2014 ◽  
Vol 71 (3) ◽  
pp. 181-186
Author(s):  
Halina Wysokińska ◽  
Katarzyna Lisowska ◽  
Katarzyna Floryanowicz-Czekalska
Keyword(s):  
Woody Plant ◽  
Storage Period ◽  
Shoot Cultures ◽  
Shoot Buds ◽  
Catalpa Ovata ◽  
Alginate Capsules ◽  
Woody Plant Medium ◽  
The Media ◽  
In Vitro Shoot Cultures

Shoot buds isolated from in vitro shoot cultures of <em>Catalpa ovata</em> G. Don were encapsulated using 3% sodium alginate with sucrose (3%) and 50 mM calcium chloride. The morphogenic response of encapsulated buds was affected by such factors, like composition of the media and the presence of growth regulators. The highest frequency of plantlet germination from encapsulated buds (70% within 4 weeks) was obtained on Woody Plant medium (WP) (Lloyd and McCown 1980) containing indole-3-butyric acid (IBA) (1 mg/l). The process was substantially inhibited by cold-storage (4<sup>o</sup>C) of encapsulated buds. In this case, the frequency response ranged from 3% to 22% dependent on storage period (28 or 42 days) and the presence of the paraffin coat covering the alginate capsules. The plantlets developed from both unstored and stored encapsulated buds of <em>C. ovata</em> were transplanted to soil and grew in pots to phenotypically normal plants.

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