scholarly journals Micropropagation of Tolumnia Orchids through Induction of Protocorm-like Bodies from Leaf Segments

HortScience ◽  
2019 ◽  
Vol 54 (7) ◽  
pp. 1230-1236 ◽  
Author(s):  
Nittaya Chookoh ◽  
Yi-Tien Chiu ◽  
Chen Chang ◽  
Wei-Hsin Hu ◽  
Ting-En Dai
Keyword(s):  
Average Rate ◽  
Survival Rates ◽  
Leaf Explants ◽  
Basal Medium ◽  
Mass Propagation ◽  
Flower Stalk ◽  
Leaf Segments ◽  
Short Time ◽  
Direct Induction

A protocol for plant regeneration via direct induction of protocorm-like bodies (PLBs) from leaf segments of Tolumnia Snow Fairy was developed as a basis for mass production. Ten-month-old, in vitro–grown donor plantlets were obtained by inducing shoots from buds on the flower stalk. Leaf segments harvested from plantlets of different heights and from expanding leaves at different positions were compared, as were two BA concentrations with 0.5 mg·L−1 NAA. The greatest rate of PLB induction (16.7%) was observed when leaf segments taken from 1- to 2-cm height plants were cultured in Murashige and Skoog (MS) basal medium supplemented with 2 mg·L−1 BA and 0.5 mg·L−1 NAA after 16 weeks of culture. When using leaf explants, only inner, expanding leaves cultured on MS basal medium supplemented with 4 mg·L−1 BA and 0.5 mg·L−1 NAA resulted in PLB induction, at an average rate of 25.5 PLBs per explant. After 16 weeks of culture, histological and scanning electron microscopy (SEM) observations revealed that PLBs originated from epidermal cells of leaf explants. PLBs of 1 to ≤2 mm in diameter continued to proliferate after 4 weeks of culture. These secondary PLBs could be produced from either whole PLBs or the upper side of PLBs. Finally, PLBs were regenerated into plantlets. After ≈14 months of culture, fully developed plants exhibiting well-developed roots and shoots were acclimatized. These plants grew well, with 1-year survival rates of nearly 73%, for plants originating as explants taken from 1- to 2-cm tall plants, and of 79%, for plants originating as explants taken from inner leaves. Some mature plants flowered 1 year after transplantation. This study presents a simple system that can provide a large number of PLBs for mass propagation in a short time that can be converted into plants and also used for the new cultivars of Tolumnia orchids.

scholarly journals Plant Regeneration from Leaf Explants of Plumbago

1970 ◽  
Vol 19 (1) ◽  
pp. 79-87 ◽  
Author(s):  
M. Gopalakrishnan ◽  
B. Janarthananm ◽  
G. Lakshmi Sai ◽  
T. Sekar
Keyword(s):  
Average Length ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Elongation ◽  
Mass Propagation ◽  
Plumbago Rosea ◽  
Half Strength ◽  
Maximum Root ◽  
Important Medicinal Plant

Leaf explants of Plumbago rosea L. an important medicinal plant inoculated on MS supplemented with 6.66 mM BAP and 2.69 mM NAA  produced numerous (105 ± 0.3) shootlets with an average length of 3.1 ± 0.0 cm. Small shootlets were transferred to shoot elongation medium supplemented with 1.11 mM BAP plus 1.44 mM GA3. The elongated shootlets transferred to half strength MS basal medium without any plant growth regulator produced 4.3 ± 0.2 rootlets per plants with average root length of 4.0 ± 0.0 cm after 25 days of culture. Rooted plantlets were transferred for hardening showed 80 - 90 per cent of plantlets success-fully established in the field. Potentially more than 50,000 plantlets could be produced within five subcultures from without callus phase obtained from leaf explant. Maximum root differentiation from leaf explants was obtained on MS supplemented with 5.38 mM IBA. The roots developed by the above method is an alternative for the controlled production of secondary metabolites. Key words:  In vitro culture, leaf explants, mass propagation, Plumbago rosea D.O.I. 10.3329/ptcb.v19i1.4989 Plant Tissue Cult. & Biotech. 19(1): 79-87, 2009 (June)

scholarly journals Mass Propagation of Nauclea diderrichii (de Willd. & Th. Dur) Merr. Seedlings through Tissue Culture Technique

2021 ◽  
Vol 31 (1) ◽  
pp. 51-60
Author(s):  
RI Oyediran ◽  
JO Afolabi ◽  
DB Olomola ◽  
FO Akanni
Keyword(s):  
Tissue Culture ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Culture Technique ◽  
Seedling Production ◽  
Mass Propagation ◽  
In Vitro Plantlets ◽  
Number Of Leaves ◽  
Nauclea Diderrichii

Nauclea diderrichii is a tree species of economic importance. However, its plantation establishment is limited by inadequate seedling production. Hence, there is ample scope of tissue culture for its mass propagation. Its in vitro plantlets development as affected by media strengths indicated that 100 % seed germination was obtained in full MS basal medium while the least (3.35 %) was from quarter-strength at 8 Weeks after inoculation (WAI). The effects of BAP and NAA assessed on the growth of its sub-cultured plantlets showed that highest number of leaves (17) and adventitious shoots (3) were obtained from MS basal medium supplemented with 0.1 mg/l BAP only. Whereas, highest shoot length (3.61 cm) and average number of roots (5/plantlet) were obtained from the same medium without hormone(s) at 8 WAI. Further sub-culturing into MS with 0.05 mg/l NAA resulted into plantlets having optimum shoot and massive root growth ready for acclimatization in 6 WAI. The plantlets were successfully acclimatized using coconuthusk/ topsoil mixture with 90 % survival. Plant Tissue Cult. & Biotech. 31(1): 51-60, 2021 (June)

scholarly journals Micropropagation of Mini Orchid Hybrid Phalaenopsis “Sogo Vivien”

2016 ◽  
Vol 1 (1) ◽  
pp. 45 ◽  
Author(s):  
Exsyupransia Mursyanti ◽  
Aziz Purwantoro ◽  
Sukarti Moeljopawiro ◽  
Endang Semiarti
Keyword(s):  
Cell Shape ◽  
Histological Analysis ◽  
The Other ◽  
Mass Propagation ◽  
Flower Stalk ◽  
Shoot Segment ◽  
Plant Body ◽  
Pot Plant ◽  
Variegated Plant

Phalaenopsis “Sogo Vivien” is an orchid hybrid with mini size plant body, and exhibits numerous beautiful pink flowers, that is ideal as ornamental pot plant. Some plants of this orchid exhibit variegated leaves that improve the beauty of the plant, not only because of the flower but also as attracted leaves. This orchid has high economical value, but mass propagation of this orchid has not established yet. An effective method to propagate both the normal and variegated plants is worth to be generated. The objective of this research was to produce a large number of P. “Sogo Vivien” plants, including the variegated plants. The method used seeds from self pollinating variegated plant, and flower stalk nodes. The seeds were sown on three various medium: VW, NP and MS, and flower stalk nodes were planted on VW + BA 10 mg l-1 + active carbon. The results showed that the best medium for in vitro culture of P. “Sogo Vivien” was NP medium, in which all seeds could grew into plantlets. Most plantlets emerged from the seeds were non variegated, only one plantlet out of 1344 seeds was variegated (0.007%). Although all emerged plantlets from flower stalk exhibited variegated leaves. Particularly, the plantlets arised from the second and third basal nodes of flower stalk showed the highest growth rate than that from the other nodes. Histological analysis showed that at 11-13 days after shoot segment plantation on NP medium, the shape of apical cells in the nodes was changed, then followed by the change of cell shape in the basal part of the nodes, produced bipolar pattern, then gradually developed into shoot. These results suggest that mass propagation could be achieved using seed culture, but to get the variegated phenotypes, the second and third nodes of flower stalk from variegated plant were the best explants to be used.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Anahi Bucchini ◽  
Laura Giamperi ◽  
Donata Ricci
Keyword(s):  
Methanol Extract ◽  
Antioxidant Activities ◽  
Leaf Explants ◽  
Basal Medium ◽  
Alternaria Solani ◽  
Antimicrobial Activities ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

scholarly journals Effect of phytohormones on shoot apex and leaf explants of Withania somnifera (Ashwagandha)

2016 ◽  
Vol 8 (1) ◽  
pp. 412-415 ◽  
Author(s):  
Archana Rani ◽  
M. Kumar ◽  
Sanjeev Kumar
Keyword(s):  
Acetic Acid ◽  
Shoot Apex ◽  
Callus Induction ◽  
Withania Somnifera ◽  
Leaf Explants ◽  
Ms Medium ◽  
Mass Propagation ◽  
Best Response ◽  
Dichlorophenoxy Acetic Acid

An efficient protocol for callus induction of Withania somnifera through in vitro culture of shoot apex and leaf explant was standardized. Of the various combinations of phytohormones evaluated, MS media supplemented with 6-furfuryl aminopurine (KIN) 0.5 mg/l + 2,4-dichlorophenoxy acetic acid (2, 4-D) 2.0 mg/l was found to be bestfor mean callus induction (86%) in leaf explants after 6 weeks of culture and in case of shoot apex expant the best response and growth of callusing was observed on MS medium supplemented with 2,4-D 1.0 mg/l + BAP 2.0 mg/l (77%).The response of callus growth increases gradually with the reductions in concentration of KIN in culturemedium of both the explants. This protocol might be used in further research for mass propagation of W. somnifera via indirect regeneration methods.

scholarly journals In vitro Multiple Shoot Regeneration from Petunia hybrida

2019 ◽  
Vol 7 (10) ◽  
pp. 1554
Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir
Keyword(s):  
Petunia Hybrida ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Length ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.

scholarly journals 117 An In Vitro Regeneration System for Basil (Ocimum basilicum L.)

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 461E-461
Author(s):  
Winthrop B. Phippen ◽  
James E. Simon
Keyword(s):  
In Vitro Regeneration ◽  
Morphological Characteristics ◽  
Leaf Explants ◽  
Basal Medium ◽  
Induction Medium ◽  
Shoot Induction ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Regeneration Procedure

A plant regeneration protocol was successfully developed for basil (O. basilicum L.). Explants from 1-month-old seedlings yielded the highest frequency of regeneration of shoots (37%) with an average number of 3.6 shoots per explant. Calli and shoot induction were initiated on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ) (4 mg/L) for ≈30 days. Shoot induction and development was achieved by refreshing the induction medium once after 14 days. The most morphogenetically responsive explants were basal leaf explants from the first fully expanded true leafs of greenhouse-grown basil seedlings. Developing shoots were then rooted on MS media in the dark without TDZ. Within 20 days, rooted plantlets were transferred and acclimatized under greenhouse conditions where they developed normal morphological characteristics. This is the first report of a successful in vitro regeneration system for basil through primary callus. The establishment of a reliable regeneration procedure is critical when developing a transformation protocol for enhancing the production of basil for insect and disease resistance and improved essential oil constituents.

scholarly journals Plant Regeneration through Protocorm-like Bodies Induced from Nodal Explants of Syngonium podophyllum ‘White Butterfly’

HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2129-2133 ◽  
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Min Deng ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Culture ◽  
Nodal Explants ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Ex Vitro ◽  
Petiole Explants

Syngonium podophyllum ‘White Butterfly’, one of the most popular ornamental foliage plants, is propagated almost exclusively through in vitro shoot culture. Ex vitro rooting, however, has been associated with severe Myrothecium leaf spot (Myrothecium roridum Tode ex Fr.). The objective of this study was to establish a method for regenerating well-rooted plantlets before ex vitro transplanting. Leaf and petiole explants were cultured on a Murashige and Skoog (MS) basal medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ), 6-benzyladenine (BA), or N-isopentenylaminopurine (2iP) with α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. Calli formed from leaf explants cultured on the basal medium supplemented with CPPU or TDZ with 2,4-D or with NAA as well as from petiole explants cultured on the medium supplemented with BA, CPPU, or TDZ with 2,4-D or NAA. The calli, however, failed to differentiate, and shoot organogenesis did not occur. Culture of nodal explants on the MS basal medium supplemented with 9.84 μm 2iP, 8.88 μm BA, 8.07 μm CPPU, or 9.08 μm TDZ with 2.26 μm 2,4-D resulted in the formation of protocorm-like bodies, adventitious shoots, and subsequently well-rooted plantlets. MS basal medium supplemented with 19.68 μm 2iP and 1.07 μm NAA resulted in the highest percentage (92.9%) of nodal explants producing protocorm-like bodies and an average of 16.9 well-rooted plantlets per nodal explant. Adventitious shoots were able to root in the initial induction medium, but better root development occurred after shoots with protocorm-like bodies were transferred onto MS basal medium supplemented with 9.84 μm 2iP and 2.69 μm NAA. Regenerated plantlets were stable and grew vigorously with 100% survival rates after ex vitro transplanting to a container substrate in a shaded greenhouse.

88 EFFECT OF VITRIFICATION PROCEDURE ON SURVIVAL RATE OF BOVINE EMBRYOS PRODUCED IN VITRO

2011 ◽  
Vol 23 (1) ◽  
pp. 149
Author(s):  
E. Y. Herrera ◽  
C. de Frutos ◽  
R. Laguna-Barraza ◽  
A. Gutierrez-Adan ◽  
D. Rizos
Keyword(s):  
Survival Rate ◽  
Survival Rates ◽  
Calf Serum ◽  
Basal Medium ◽  
Bovine Embryos ◽  
Oviduct Fluid ◽  
Bovine Oocytes ◽  
Cryopreservation Method ◽  
Hatching Rates

Vitrification as a cryopreservation method has many advantages compared with slow freezing. Many variables in the vitrification process exists that influence the survival rates of vitrified oocytes and embryos. These include the cryoprotectants (type, concentration, and duration of exposure), the temperature of the vitrification solution at exposure, the device used for vitrification, and the quality and developmental stage of embryos. It is worthwhile to mention that vitrification protocols successfully used in bovine oocytes and embryos have been used also with human oocytes and embryos. Vitrification is relatively simple, requires no freezing equipment, and relies on the placement of the embryos in a very small volume of vitrification medium that must be cooled at extreme rates not obtainable in regular enclosed straws. The aim of the present study was to evaluate the efficiency of 4 different vitrification protocols on the survival rate of in vitro produced (IVP) bovine embryos. Blastocysts were produced by a standard IVP procedure following in vitro maturation, fertilization, and culture in synthetic oviduct fluid supplemented with 5% fetal calf serum (FCS). On Day 7 (Day of IVF = Day 0), a total of 297 blastocysts were vitrified using (i) the open pulled straw (OPS) in 20% DMSO and 20% ethylene glycol (EG) in a basal medium of TCM-199 with HEPES supplemented with 20% FCS; (ii) the modified OPS, in 20% DMSO, 20% EG, and 0.5 M sucrose in a basal medium of phosphate buffer saline (PBS) supplemented with 20% FCS; (iii) the cryoloop, in 15% DMSO, 15% EG, 10 mg mL–1 Ficoll 70, and 0.65 M sucrose in a basal medium of PBS supplemented with 20% FCS; and (iv) in 0.25 straws in 20% glycerol, 20% EG, 0.3 M sucrose, 3% polyethylene glycol, and 0.3 M xylose in a basal medium of PBS. After warming, embryos were placed in culture for additional 24 h. Re-expansion and hatching rates were measured at 2 and 24 h after warming. Data were analysed by 1-way ANOVA. At 2 h post-warming, the re-expansion of blastocysts vitrified with cryoloop was significantly higher compared with OPS, modified OPS, and the 0.25 straw methods (54.08 ± 15.53 v. 10.40 ± 3.00, 22.67 ± 9.20, and 8.82 ± 2.15, respectively; P ≤ 0.028). At 24 h post-warming, only embryos from cryoloop and modified OPS were still alive with a survival rate of embryos vitrified with cryoloop significantly higher than that of those vitrified with modified OPS (48.45 ± 17.56 v. 3.75 ± 3.75, respectively; P ≤ 0.007). Hatching rates at 24 h post-warming were not different between cryoloop and modified OPS groups (5.63 ± 4.40 and 1.25 ± 1.25, respectively). These results clearly demonstrate that embryo cryotolerance is affected by the method used for cryopreservation. Moreover, cryoloop vitrification was found to be more effective than OPS and 0.25 straw methods for the cryopreservation of bovine embryos.

Sign in / Sign up

Export Citation Format

Share Document