scholarly journals Development of An Efficient Protocol for Rapid Propagation of Curculigo orchioides Using Leaf Explant Culture

2021 ◽  
Vol 14 (4) ◽  
pp. 1514-1519
Author(s):  
Tran Thi Linh Giang
Keyword(s):  
Leaf Explants ◽  
Basal Medium ◽  
Culture Method ◽  
Explant Culture ◽  
Regeneration Rate ◽  
Curculigo Orchioides ◽  
Rapid Propagation ◽  
Best Response ◽  
Shoot Number

Curculigo orchioides is one of the most common medicinal plants used by diverse cultures and tribal groups. The roots of the plant are used medicinally in Asian countries. Curculigo orchioides have the ability to regenerate through seeds and tubers, but the regeneration rate is low. Plant tissue culture method was believed to have potential for rapid multiplication of this medicinal plant. An efficient protocol for rapid propagation of Curculigo orchioides, of the family Amaryllidaceae, was developed using leaf explants culture. The leaf explants (1 cm x 1 cm squares) cultured on Murashige and Skoog (MS) basal medium were supplemented with various concentrations and combinations of auxins and cytokinins with temperature 25 ± 2°C, relative humidity 85-90% and photoperiod of 12 hours light (2000-3000 lux). Callus induction was obtained within 4 weeks, 2,4-D at 3 mg/l formed profuse callus and the degree was found to be the highest (+++) among all the treatments. The best response to shoot induction, with maximum shoot number 5.33 (mean number of shoots per explant) was obtained using 1.0 mg/l 6-benzyl aminopurine (BAP) in combination with 1.0 mg/l Kinetin. In vitro shoots were induced for rooting on 0.5 mg/l of NAA supplemented medium. In order for seedlings propagated in vitro to adapt to natural conditions, plants were growned on a substrate coir: husk ash: sand (with the ratio of 0.5: 0.5: 1) in a greenhouse (humidity: 70%, temperature: 28-300C) gave 88.33% survival rate after 8 weeks of culture. With the results received, this is an effective approach to propagating Curculigo orchioides.

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 Regeneration of Chlorophytum amaniense ‘Fire Flash’ Through Indirect Shoot Organogenesis

HortScience ◽  
2011 ◽  
Vol 46 (3) ◽  
pp. 466-469
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Callus Formation ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Culture Method ◽  
Shoot Formation ◽  
Photon Flux ◽  
Naphthalene Acetic Acid ◽  
Foliage Plant

Chlorophytum amaniense Engl. ‘Fire Flash’ is a popular exotic ornamental foliage plant as a result of its unique coral-colored midribs and petioles and tolerance to interior low light levels. Currently, demand for propagative materials exceeds the availability of seeds. This study was intended to develop an in vitro culture method for rapid propagation of this cultivar. Leaf and sprouted seed explants were cultured on a Murashige and Skoog basal medium supplemented with different cytokinins with 1.1 μM α-naphthalene acetic acid (NAA) or 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf explants showed poor responses in callus production and no adventitious shoots were obtained. Callus formation frequencies from sprouted seeds were 71% and 85% when induced by 9.8 μM N6-(2-isopentyl) adenine (2iP) with 1.1 μM NAA and 9.1 μM N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with 1.1 μM NAA, respectively. Adventitious shoots occurred after the induced calluses were subcultured on the same concentrations of TDZ or 2iP with NAA. Shoot formation frequencies from calluses cultured on TDZ with NAA and 2iP with NAA were 92% and 85%, and the corresponding mean shoot numbers were 37 and 31 per piece of callus (1 cm3), respectively. Adventitious shoots rooted at 100% after transferring to the basal medium containing 4.4 μM 6-benzylaminopurine (BA) with 2.7 μM NAA. Plantlets, after transplanting to a soilless substrate were easily acclimatized in a shaded greenhouse under a photosynthetic photon flux (PPF) density of 200 μmol·m−2·s−1. Regenerated plants grew vigorously without undesirable basal branching or distorted leaves. This newly established regeneration method can provide the foliage plant industry with a means for rapidly propagating ‘Fire Flash’ liners in a year-round fashion.

scholarly journals Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

2022 ◽  
Vol 0 (0) ◽  
Author(s):  
Hamid Reza SABAGHI ◽  
Gholamreza SHARIFI-SIRCHI ◽  
Pejman AZADI ◽  
Mohammad Hossein AZIMI
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Dianthus Caryophyllus ◽  
Casein Hydrolysate ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Rate ◽  
Shoot Number

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaves of Formosan Sweetgum (Liquidambar formosana L.)

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 721-723 ◽  
Author(s):  
L. Xu ◽  
G.F. Liu ◽  
M.Z. Bao
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
High Regeneration

Plantlets were regenerated from in vitro-grown leaf explants of five genotypes of Liquidambar formosana on WPM basal medium supplemented with different concentrations of TDZ and NAA. With the addition of 0.27 μm NAA, regeneration efficiency was increased by 2- to 4-fold over that with TDZ alone. Lower concentrations of TDZ (0.45–2.27 μm) were beneficial for regenerating shoot clusters. Four genotypes (P2, P6, P9, and P11) showed high regeneration rates (up to 90%), whereas genotype P13 showed a low capability for shoot regeneration on all media tested (<35%). For all five genotypes, the optimum medium for inducing adventitious shoots was WPM supplemented with 1.14 μm TDZ and 0.27 μm NAA, on which regeneration rate ranged from 72.6% to 89.5% and adventitious shoot clusters per regenerating leaf explant ranged from 2.63 to 3.11 in four genotypes (P2, P6, P9, and P11), while for P13, the regeneration rate and number of shoot clusters per regenerating explant were 23% and 1.39, respectively. Transfer of shoot clusters to WPM basal medium containing 0.54 μm NAA, 2.22 μm BA, and 1.44 μm GA3, resulted in shoot elongation. All the elongated shoots were rooted on WPM supplemented with 9.84 μm IBA, and plantlets were transplanted to soil successfully. Chemical names used: 6-benzyladenine (BA), gibberellic acid (GA3), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), plant growth regulator (PGR), thidiazuron (TDZ), woody plant medium (WPM).

scholarly journals Mesenchymal Stem/Stromal Cells from Discarded Neonatal Sternal Tissue: In Vitro Characterization and Angiogenic Properties

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Shuyun Wang ◽  
Lakshmi Mundada ◽  
Eric Colomb ◽  
Richard G. Ohye ◽  
Ming-Sing Si
Keyword(s):  
Bone Marrow ◽  
Cardiac Surgery ◽  
Stromal Cells ◽  
Umbilical Vein ◽  
Culture Method ◽  
Tissue Perfusion ◽  
Explant Culture ◽  
Critical Congenital Heart Disease ◽  
Surface Marker Expression

Autologous and nonautologous bone marrow mesenchymal stem/stromal cells (MSCs) are being evaluated as proangiogenic agents for ischemic and vascular disease in adults but not in children. A significant number of newborns and infants with critical congenital heart disease who undergo cardiac surgery already have or are at risk of developing conditions related to inadequate tissue perfusion. During neonatal cardiac surgery, a small amount of sternal tissue is usually discarded. Here we demonstrate that MSCs can be isolated from human neonatal sternal tissue using a nonenzymatic explant culture method. Neonatal sternal bone MSCs (sbMSCs) were clonogenic, had a surface marker expression profile that was characteristic of bone marrow MSCs, were multipotent, and expressed pluripotency-related genes at low levels. Neonatal sbMSCs also demonstrated in vitro proangiogenic properties. Sternal bone MSCs cooperated with human umbilical vein endothelial cells (HUVECs) to form 3D networks and tubes in vitro. Conditioned media from sbMSCs cultured in hypoxia also promoted HUVEC survival and migration. Given the neonatal source, ease of isolation, and proangiogenic properties, sbMSCs may have relevance to therapeutic applications.

scholarly journals Micropropagation of Tarenaya rosea (Cleomaceae) from leaf explants

Rodriguésia ◽  
2021 ◽  
Vol 72 ◽  
Author(s):  
Claudia Simões-Gurgel ◽  
Tatiana Carvalho de Castro ◽  
Cátia Henriques Callado ◽  
Lívia da Silva Cordeiro ◽  
Norma Albarello
Keyword(s):  
Large Scale ◽  
Plant Conservation ◽  
Leaf Explants ◽  
Plant Production ◽  
Anthocyanin Content ◽  
Alternative Source ◽  
Regeneration Rate ◽  
Culture Techniques ◽  
Regenerated Plants

Abstract In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.

scholarly journals In vitro Propagation of Vanda testacea (Lindl.) Reichb.f. – A Rare Orchid of High Medicinal Value

2010 ◽  
Vol 19 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Saranjeet Kaur ◽  
K.K. Bhutani
Keyword(s):  
Leaf Explants ◽  
Subsequent Development ◽  
Individual Treatment ◽  
Chemical Stimulus ◽  
Medicinal Value ◽  
Best Response ◽  
High Regeneration ◽  
Regeneration Response ◽  
High Regeneration Frequency

Foliar explants of Vanda testacea (Lindl.) Reichb. f. were cultured on Mitra (M) medium with 1.0 mg/l BAP, Kn  each and 1.0 mg/l NAA individually and in combination for initiation of regeneration response, proliferation of regenerants and subsequent development of plantlets. Juvenility of the tissues and chemical stimulus were important factors in initiating the regeneration response in the explants. The relatively older leaf explants (>1cm in length) remained recalcitrant to regeneration the representing younger ones (<1cm in length) responded to certain chemical regimes. BAP, Kn individually in the medium should direct PLB regeneration whereas when used with NAA, the explants showed callus proliferation and further differentiated into PLBs. An individual treatment with NAA (1.0 mg/l) impaired the response frequency and delayed further morphogenetic processes leading to plantlet development. The best response in the explants (in terms of high regeneration frequency, early initiation, PLB proliferations, and plantlet development) was observed in 1.0 mg/l BAP alone/with 1.0 mg/l NAA + activated charcoal. Plantlets were transferred to pots containing epiphytic compost (1 charcoal : 1 brick pices : 1 bats). Nearly 75% of plantlets survival was recorded.  Key words: In vitro, Orchid, Vanda testacea, Micropropagation, Protocorm-like bodies, callus D.O.I. 10.3329/ptcb.v19i1.4077 Plant Tissue Cult. & Biotech. 19(1): 1-7, 2009 (June) 

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.

REGENERATION OF TREMA ORIENTALIS (BLUME) LINN.: EFFECT OF GROWTH REGULATORS, CULTURE CONDITIONS, AND AGE AND SOURCE OF EXPLANTS

1995 ◽  
Vol 43 (4) ◽  
pp. 391-395 ◽  
Author(s):  
G.R. Rout ◽  
S. Samantaray ◽  
P. Das
Keyword(s):  
Growth Regulators ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Mature Trees ◽  
Regeneration Rate ◽  
Bud Regeneration ◽  
Shoot Bud ◽  
Shoot Bud Regeneration ◽  
Trema Orientalis

Optimal conditions for high frequency shoot bud regeneration from leaf callus of Trema orientalis (Blume) Linn. were studied. The regeneration rate was controlled by the growth regulators, the age and the source of the explants, and the illumination conditions. Irrespective of illumination conditions, shoot bud regeneration was achieved only in media containing benzyladenine (BA) + α-naphthaleneacetic acid (NAA) combinations, with the best results being obtained in the presence of 2.5 mg/1 BA and 0.25–0.5 mg/1 NAA. The morphogenic response was less frequent in the calluses derived from leaf explants of the mature trees compared to those of the in vitro-grown seedlings. The rate of shoot bud regeneration was more pronounced in the cultures maintained for 4 weeks in the light (16-h photoperiod) than the cultures incubated in the dark. Regenerated shoots were rooted on the medium containing 1/2 strength basal Murashige and Skoog (MS) salts supplemented with 0.01 mg/1 NAA or indole-3-butyric acid (IBA). The rooted plantlets were established in the greenhouse.

scholarly journals TDZ Plays Key Role in Shoot Regeneration from Different Explants of Picrorhiza kurroa: An Endangered Medicinal Herb of Western Himalayas

2019 ◽  
pp. 1-9 ◽  
Author(s):  
Vanita Patial ◽  
Amita Bhattacharya
Keyword(s):  
Natural Habitat ◽  
Basal Medium ◽  
In Vitro Rooting ◽  
Western Himalayas ◽  
Picrorhiza Kurroa ◽  
Regeneration Potential ◽  
Root Explants ◽  
Shoot Number ◽  
Endangered Medicinal Herb

Picrorhiza kurroa plants were collected from its natural habitat. In vitro plants were raised from the leaves of high yielding collection screened in an earlier study. Leaves, roots and internodal segments were cultured for 15 days. The effect of thidiazuron (1-phenyl-3-(1, 2, 3- thiadiazol-5-yl) urea; TDZ) pretreatment for 15 days on regeneration potential of different explants viz. leaves, roots and internodes of Picrorhiza kurroa was studied. Regeneration potential varied significantly with the type of explant. Regeneration response of 100% with 46.25 shoots per explant was obtained in leaf segments of 2.0 cm length pretreated with 0.5 µM TDZ for 15 days and then transferred to 2.32 µM kinetin (Kn) containing MS basal medium. In case of root explants maximum shoot number (17.12) was obtained on 0.5 µM TDZ pretreated for 15 days and then to 3.64 µM Kn. Maximum shoots per explants (12.33) were obtained in case of internodes pretreated with 0.5 µM TDZ for 15 days and transferred to 1.16 µM Kn. Regenerated shoots from different explants developed in vitro rooting on MS basal medium within 7-8 days. Conclusively, an efficient and repeatable protocol for rapid regeneration from different explants and in vitro rooting has been developed in P. kurroa which can be effectively used for its conservation.

Sign in / Sign up

Export Citation Format

Share Document