Establishment of in vitro aseptic culture of Philodendron xanadu Croat

A micropropagation protocol for an endangered slipper orchid species, Cypripedium formosanum Hayata, through axillary buds from adult plants has been developed. The season of explant collection is crucial for the initial success of an aseptic culture. Explants collected in the middle of January gave the highest percentage of explant survival (54.2%) and shoot-forming percentage (41.7%). Of the two cytokinins tested, N6-benzyladenine (BA) was found to be superior to thidiazuron for normal shoot formation. The optimum result was obtained in quarter-strength Murashige and Skoog medium containing 22.2 or 44.4 μM BA in which the cultures produced 6.3 and 7.1 shoots per explant with 10.6 to 11.7 mm average length after 90 d of culture. Regenerated shoots rooted for 60 d in the basal medium with 1 g·L−1 activated charcoal and 20 g·L−1 potato homogenate were ready for growth in pots. This is the first report on shoot multiplication in vitro from mature plants of Cypripedium that provides a reliable method for propagating the selected elites.

Download Full-text

Method of growing aseptic culture of zingiber in vitro

Visnyk agrarnoi nauky ◽

10.31073/agrovisnyk201709-07 ◽

2017 ◽

Vol 95 (9) ◽

pp. 40-43

Author(s):

N. Beh ◽

M. Kotsar ◽

M. Roik

Keyword(s):

Aseptic Culture

Download Full-text

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.

Download Full-text

Shoot Regeneration in vitro From Native Australian Fruit-Bearing Trees - Quandong and Plum Bush

Australian Journal of Botany ◽

10.1071/bt9800405 ◽

1980 ◽

Vol 28 (4) ◽

pp. 405 ◽

Cited By ~ 7

Author(s):

M Barlass ◽

WJR Grant ◽

KGM Skene

Keyword(s):

Shoot Growth ◽

Adventitious Shoot ◽

Indolebutyric Acid ◽

Axillary Shoots ◽

Aseptic Culture ◽

Aerial Parts ◽

Edible Fruit ◽

Semiarid Areas ◽

Regeneration In Vitro

Explants from quandong (Santalum acuminatum) and plum bush ( S. lanceolatum), native Australian trees, regenerated shoots in aseptic culture. In the presence of cytokinin. shoots were produced in quantity from all cultured aerial parts of quandong seedlings. Individually cultured nodes of I-yr-old plum bush and quandong plants proliferated axillary shoots, while internodal pieces produced adventitious shoot growth. Shoots were also regenerated from explants of a 5-yr-old mature quandong tree. Excised in vitro-grown shoots produced roots in culture on a medium containing indolebutyric acid at pH 4 following shoot etiolation. These results offer the promise of clonal propagation in species in which conventional vegetative propagation by cuttings has not been reported, and will provide a useful adjunct to a quandong breeding program to develop edible fruit and nuts in semiarid areas.

Download Full-text

In vitro propagation and medium-term conservation of autochthonous plum cultivar 'Crvena Ranka'

Acta agriculturae Serbica ◽

10.5937/aaser2050141v ◽

2020 ◽

Vol 25 (50) ◽

pp. 141-147

Author(s):

Tatjana Vujović ◽

Darko Jevremović ◽

Tatjana Marjanović ◽

Ivana Glišić

Keyword(s):

In Vitro Propagation ◽

Sustainable Use ◽

Shoot Multiplication ◽

Growth Conditions ◽

High Survival ◽

Indigenous Species ◽

Medium Term ◽

Aseptic Culture ◽

In Vitro Shoots

In vitro strategies for the propagation and conservation of indigenous species contribute to the sustainable use of plant diversity and are essential for breeding programs as well. In this study, we established an efficient protocol for the micropropagation of autochthonous plum 'Crvena Ranka' and examined the survival and regrowth capacity of in vitro shoots after 3, 6 and 9 months of cold storage (CS) at +5 oC in total darkness. Aseptic culture was established on the Murashige and Skoog medium containing 2 mg l-1 BA, 0.5 mg l-1 IBA and 0.1 mg l-1 GA3 (leaf rosette initiation being 68.8%). During in vitro propagation on the medium of constant hormonal composition, a significant increase in the multiplication index was observed in the third subculture, whereupon it was mainly stable until the fifth subculture. The effect of BA concentration and/or type of auxins (IBA or NAA) on multiplication parameters, as well as on fresh and dry weights of shoots was evaluated. BA at 1 mg l-1 in combination with NAA significantly increased shoot multiplication parameters. The effect of auxins on rooting parameters was monitored as well. Shoots cultured on the medium supplemented with NAA also displayed higher rooting ability (60%), in comparison with those grown on the medium containing IBA at the same concentration (20%). In vitro shoots can be conserved over the medium term under CS conditions up to six months. High survival was achieved after three (94%) and six months (82.5%), while severe signs of necrosis (100%) were noticed after nine months of conservation. Shoots subcultured under standard growth conditions after CS promptly regained their morphology although their capacity for multiplication and rooting was slightly lower than that of non-cold-stored shoots.

Download Full-text

901 PB 538 STUDIES ON PROPAGATION OF GLOBE ARTICHOKE THROUGH TISSUE CULTURE TECHNIQUE

HortScience ◽

10.21273/hortsci.29.5.563b ◽

1994 ◽

Vol 29 (5) ◽

pp. 563b-563 ◽

Cited By ~ 1

Author(s):

Kh. A. Okasha ◽

M. E. Ragab

Keyword(s):

Tissue Culture ◽

Culture Technique ◽

Multiplication Rate ◽

Globe Artichoke ◽

Ms Medium ◽

Free Living ◽

Survival Percentage ◽

Aseptic Culture ◽

Low Rate

The aim of this study was to establish an effective method of micropropagation of globe artichoke from shoot-hp. This study involved establishment of an aseptic culture. multiplication of proliferated shook, rooting of these shook in vitro and adaptation of plantlets for free living Highest survival percentage with no contamination was achieved after sterilizing the explants with 70% ethanol (5 Sec.) + 1.5 % sodium hypochlorite for 20 min. The bast results of preventing browning of explants were obtained with 100 mg/1 ascorbic acid. The highest proliferated shook were obtained when shoot tip with 4 mm length (taken in mid March) were cultured on MS medium with 10mg/1 Kin. + 0.5 mg/l IAA. The highest multiplication rate was obtained when recultured on MS medium supplemented with 5 mg/1 Kin. + 0.5 mg/1 IAA. Multiplication rate gradually increased with increasing number of subculturing till Me third subculture but subsequent subculture (4th subculture) had a low rate.

Download Full-text

OPTIMAL GROWTH, RAPID ACCLIMATIZATION AND SHIPPING OF CATTLEYA ORCHID IN SINGLE USE SEALED MEMBRANE VESSELS

HortScience ◽

10.21273/hortsci.28.4.263e ◽

1993 ◽

Vol 28 (4) ◽

pp. 263E-263 ◽

Cited By ~ 1

Author(s):

Laura A. Dellevigne ◽

Jeffrey W. Adelberg ◽

Peter Vergano

Keyword(s):

Three Dimensional ◽

Side Wall ◽

Optimal Growth ◽

Liquid Media ◽

Polypropylene Membrane ◽

Aseptic Culture ◽

Temperature Regimes ◽

Ex Vitro Growth ◽

Single Use

Three-dimensional polypropylene enclosures have been fabricated for the in vitro culture and ex vitro growth of Cattleya orchid propagules. The enclosures consist of: 1) microporous polypropylene membrane for nutrient transfer between liquid media and the growing tissue. 2) molded polypropylene side wall sized for growth of Cattleya orchid plants and flanged to allow heat seals with membranes, and 3) polypropylene membrane(s) top member for light and gaseous transmission. Three commercial clones of Cattleya have been sealed into these enclosures and grown for eight months on unmended MS medium. Contaminated liquid media was effectively isolated from the propagules within the sealed enclosures, and following a bleach treatment with sterile rinses, propagules were returned to aseptic culture. Greenhouse growth of plant tissues in these enclosures will be discussed. Optimization for growth of Cattleya has begun with studies of gas, light and temperature regimes within the sealed enclosures and a comparison of growth on two different nutrient formulations.

Download Full-text

Propagation of edible honeysuckle (Lonicera edulis Turcz) in in vitro conditions

Agricultural science and practice ◽

10.15407/agrisp5.02.018 ◽

2018 ◽

Vol 5 (2) ◽

pp. 18-26

Author(s):

Ya. Zapolsky ◽

T. Medvedeva ◽

T. Natalchuk ◽

M. Bublyk

Keyword(s):

Culture Medium ◽

Positive Impact ◽

Mercury Chloride ◽

Free Medium ◽

Aseptic Culture ◽

Effi Ciency ◽

Comparison Results ◽

The Impact

Aim. To propagate edible honeysuckle (Lonicera edulis Turcz) in in vitro conditions; to study the impact of sterilization agents on honeysuckle explants; to investigate the impact of the culture medium composition on the coeffi cient of propagation and rooting; to study the capability to adapt to in vivo conditions. Methods. Laboratory, mathematical, estimation and comparison. Results. The impact of sterilizing substances on obtain- ing the aseptic culture of edible honeysuckle in in vivo conditions was studied. The experiments were con- ducted on the following species: Alicia, Spokusa, Chaika, Nimfa, Doch Velikana, Karina. Lisoformin 3000 and mercury chloride were used as sterilizing agents. In the variant with Lisoformin 3000 it was studied in three exposures – 5, 7, and 10 minutes. In terms of explant regeneration effi ciency after sterilization with Lisofor- min 3000, three groups of edible honeysuckle species were isolated: 1 – with high regeneration capacity (94– 96 %) – Alicia, Karina and Spokusa; 2 – medium capacity (86–87 %) – Chaika and Doch Velikana, 3 – low capacity (80 %) – Nimfa. The experiments aimed at studying the impact of culture medium components on the propagation effi ciency determined the increase in the latter in case of rotating media with different quantitative and qualitative composition. Permanent application of uniform media leads to a sharp decrease in the prolif- eration coeffi cient in all the investigated species. Both hormone-free medium and the medium with growth regulators are effi cient for rooting. High indices of rooting were achieved in both variants. The use of auxins promoted the formation of a larger amount of plant roots (from 3.09 in Spokusa to 4.21 in Alicia) which in its turn impacted the survivability of plants in in vivo conditions. Conclusions. It was established that Lisoformin 3000 in the concentration of 3 % and at the exposure duration of 5 min ensured optimal effi ciency of steriliza- tion and regeneration of edible honeysuckle explants and did not decrease their propagation coeffi cients. With corresponding concentrations and sterilization duration, this preparation may be recommended for obtaining the aseptic culture of honeysuckle. It was demonstrated that the rotation of media, rich and poor in macro- and microsalts was effi cient for obtaining high indices of proliferation: the plants had a larger amount of tillering even in case of using not high concentrations of cytokinin. The introduction of rhizogenesis inducer, IBA, (1 mg/l) into the culture medium did not increase the percentage of rooted plants compared to hormone-free medium, but stimulated the formation of a larger amount of roots, which had further positive impact on the adaptation properties.

Download Full-text

Optimization Studies of Culture Media for In-Vitro Clonal Micropropagation of New Grape Varieties

KnE Life Sciences ◽

10.18502/kls.v0i0.9013 ◽

2021 ◽

Author(s):

Abdulmalik Batukaev ◽

Eliza Sobralieva ◽

Diana Palaeva

Keyword(s):

Culture Medium ◽

Culture Media ◽

In Vitro Regeneration ◽

Mineral Salts ◽

Additional Increase ◽

Shoot Bud ◽

Clonal Micropropagation ◽

Aseptic Culture ◽

Grape Varieties

This article describes the effect of Gautheret, White, Heller and Murashige & Skoog mineral salts during in-vitro clonal micropropagation of new grape varieties. The optimal mineral compositions of the culture medium that support the in-vitro regeneration of isolated grape explants were identified. The grapes that were studied were the Bart and Augustine varieties. Primary grape explants were cultivated for 30 days in a non-transplanted culture. Increased regenerative activity was observed in the Murashige & Skoog and White media. Increased haemogenesis occurred and shoots regenerated. The addition of cytokinin 6-BAP to the medium for obtaining aseptic culture led to an increase in the frequency of shoot-bud production by 5 to 6 times, depending on the type of medium. Combining 6-BAP with the auxin NAA provided an additional increase in the frequency of shoot-bud production, but to a lesser extent. Adding growth regulators to the culture medium also reduced the frequency of explant necrosis. Keywords: grapes, mineral salts, culture medium, microclonal propagation, in-vitro, cytokinins, auxins

Download Full-text