In vitro Regeneration of Picralima nitida (Stapf). T. Durand & H. using Zygotic Embryo

Disinfected mature seed embryos of Picralima nitida, were cultured in MS medium supplemented with different concentrations and combinations of 2,4-D, BAP and NAA to determine an efficient protocol for in vitro propagation. Nine culture media made of combination of different components were used in a factorial design with three replications. Results showed up to 80 ± 4% disinfection rate with combination of triton x- 100 (0.2%) and sodium hypochlorite (30%). Embryo germination was highest on control medium. Rooting was higher (2±1 roots per embryo) after 4 weeks on control medium and on BAP supplemented medium at 0.8 μM while the longest root (1.5±0.5 cm) was observed on 2,4-D supplemented medium at 1.8 μM. Black soil was suitable for leaf formation (4 ± 2 leaves) and shoot elongation (2±1 cm) after 8 weeks in acclimatisation. These results show efficient disinfection, regeneration and acclimatisation of Picralima nitida. Plant Tissue Cult. & Biotech. 31(2): 143-151, 2021 (December)

Download Full-text

Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

Polysaccharides ◽

10.3390/polysaccharides2020032 ◽

2021 ◽

Vol 2 (2) ◽

pp. 538-553

Author(s):

Natacha Coelho ◽

Alexandra Filipe ◽

Bruno Medronho ◽

Solange Magalhães ◽

Carla Vitorino ◽

...

Keyword(s):

In Vitro Culture ◽

Culture Medium ◽

Culture Media ◽

Average Pore Size ◽

Physiological Responses ◽

Gum Arabic ◽

Shoot Elongation ◽

Hydroxyethyl Cellulose ◽

Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

Download Full-text

Types and concentrations of cytokinins in the micropropagation of Anthurium maricense

Revista Agro mbiente On-line ◽

10.18227/1982-8470ragro.v12i2.4671 ◽

2018 ◽

Vol 12 (2) ◽

pp. 117

Author(s):

Cecília Moreira Serafim ◽

Arlene Santisteban Campos ◽

Priscila Bezerra Dos Santos Melo ◽

Ana Cecília Ribeiro de Castro ◽

Ana Cristina Portugal Pinto de Carvalho

Keyword(s):

Light Intensity ◽

Culture Medium ◽

Culture Media ◽

In Vitro Regeneration ◽

Nodal Segments ◽

Nodal Segment ◽

Viable Option ◽

Growth Room ◽

In Vitro Induction

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.

Download Full-text

Essential factors for in vitro regeneration of rose and a protocol for plant regeneration from leaves

Horticultural Science ◽

10.17221/12/2017-hortsci ◽

2018 ◽

Vol 45 (No. 2) ◽

pp. 83-91

Author(s):

Anber Mahmoud Ahmed Hassanein ◽

Inas Mohamed Ali Mahmoud

Keyword(s):

Indole Acetic Acid ◽

Culture Medium ◽

Callus Formation ◽

In Vitro Regeneration ◽

Shoot Elongation ◽

Favorable Effect ◽

Indole Butyric Acid ◽

Leaf Discs ◽

Better Than

In vitro propagation of Rosa hybrida, L. cv. ‘Eiffel Tower’ was improved by the addition of thidiazuron (TDZ) and silver nitrate (AgNo3) to the culture medium. The combination of auxin and cytokinins was indispensable for inducing response from leaf discs. Maintaining cultures under dark was better than light for callus formation and quality. The source of explants was vital in the regeneration process wherein situ explants produced callus while, in vitro explants regenerated somatic embryos and shoots. Gibberellic acid (GA3) had a favorable effect where in vitro explants showed somatic embryogenesis with no shoots on media containing TDZ however, 37% of explants regenerated shoots directly on medium containing GA3. The presence of benzyl adenine (BA) was essential for shoot elongation, and indole butyric acid (IBA) was better than indole acetic acid (IAA) for rooting. The optimum conditions produced rooted plants from leaf discs within ten weeks. The reported results clarify factors controlling in vitro regeneration of R. hybrida, and provide a rapid protocol allowing further improvements of rose.

Download Full-text

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

Canadian Journal of Plant Science ◽

10.4141/p05-055 ◽

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

Download Full-text

In Vitro Regeneration of Phyllanthus amarus Schum. and Thonn.: An Important Medicinal Plant

Our Nature ◽

10.3126/on.v7i1.2557 ◽

1970 ◽

Vol 7 (1) ◽

pp. 110-115 ◽

Cited By ~ 3

Author(s):

A. Sen ◽

M.M. Sharma ◽

D. Grover ◽

A. Batra

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Shoot Elongation ◽

Nodal Segments ◽

Nodal Segment ◽

Phyllanthus Amarus ◽

Regenerated Plants ◽

Half Strength ◽

Important Medicinal Plant

An efficient in vitro plant regeneration protocol was developed for the medicinally potent plant species Phyllanthus amarus Schum. and Thonn. (Euphorbiaceae) using nodal segment as explant. Maximum multiplication of shoots (15.275±0.96) was achieved on Murashige and Skoog’s medium supplemented with BAP (0.5 mg/l) after 3-4 weeks of inoculation. The shoots were separated from cluster and subcultured for their elongation on the same medium. In vitro flowering was also observed on the elongated shoots after 3–4 weeks of sub culturing on the shoot elongation medium. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l). Regenerated plants were successfully hardened and acclimatized, 80 % of plantlets survived well under natural conditions after transplantation.Key words: In vitro regeneration, multiple shoots, nodal segments, Phyllanthus amarusDOI: 10.3126/on.v7i1.2557Our Nature (2009) 7:110-115

Download Full-text

Coconut Tissue Culture: The Indian Initiatives, Experiences and Achievements

CORD ◽

10.37833/cord.v33i2.48 ◽

2017 ◽

Vol 33 (2) ◽

pp. 11

Author(s):

Anitha Karun

Keyword(s):

Tissue Culture ◽

Somatic Embryos ◽

Culture Media ◽

Zygotic Embryo ◽

Shoot Meristem ◽

Direct Organogenesis ◽

Zygotic Embryos ◽

Immature Inflorescence ◽

High Yielding Varieties

Coconut is one of the principal crops of India cultivated in over 35 districts mainly in the southern states. The productivity of the crop is declining in many of the traditionally cultivated regions owing to ageing plantations as well as biotic and abiotic stresses. These plantations are to be replanted with high yielding varieties/hybrids for which adequate quantity of quality planting material is not available. Even though tissue culture research was initiated in many laboratories in the country, the work was eventually phased out in most of the laboratories for want of a repeatable protocol. At ICAR-CPCRI, coconut tissue culture programs have been continuing for the past three decades. The attempts made include experimentation with different explants viz., immature inflorescence, plumular tissues, mature palm shoot meristem, ovary and anthers and different culture media supplemented with varying levels and types of hormones. Some of the successful protocols developed at the Institute include coconut zygotic embryo culture for collection and exchange of germplasm, cryopreservation and retrieval of zygotic embryos and pollen and plantlet regeneration from plumular tissues. Even though ICAR-CPCRI has succeeded in obtaining plantlets via direct organogenesis from inflorescence explants, the absence of friable calli formation from explants, the low rate of somatic embryo formation, large number of cultures turning to abnormal shoot development, non conversion of somatic embryos into plantlets, and formation of abnormal somatic embryos remain the major bottlenecks. Gene expression studies are being currently undertaken to decipher the molecular basis of in vitro recalcitrance.

Download Full-text

In vitro Regeneration and Callogenesis of Libidibia ferrea

Journal of Experimental Agriculture International ◽

10.9734/jeai/2020/v42i430495 ◽

2020 ◽

pp. 14-24

Author(s):

Daniel da Silva ◽

Angela Maria Imakawa ◽

Kamylla Rosas Vieira Guedes ◽

Flávio Mauro Souza Bruno ◽

Paulo de Tarso Barbosa Sampaio

Keyword(s):

Culture Media ◽

In Vitro Regeneration ◽

Shoot Multiplication ◽

Light Sources ◽

Multiplication Rate ◽

Medicinal Species ◽

Efficient System ◽

Blue Led ◽

Friable Callus

Libidibia ferrea (Fabaceae) is a valuable medicinal species in the Amazon, but as it is a protected plant, collection from natural populations is forbidden. Therefore, establishing an efficient system for in vitro regeneration and to improve callogenesis of this species is desirable. To determine the optimal nutritional factors needed for shoot multiplication and callus induction, different culture media, plant growth regulators and LED light sources were tested. The data were subjected to analysis of variance (ANOVA) and means compared by Tukey’s test at p < 0.05. We observe that explants inoculated in the Murashige and Skoog (MS) media with 0.05 mg L-1 of 6-benzilaminopurine (BAP) and cultivated under red-blue LED induced the highest number of shoots (3.67), number of buds (3.13), multiplication rate (15.67) and shoots length (22.03 mm) when compared with other treatments. MS and B5 media supplemented with 2.21 and 4.42 mg L-1 of 2,4-D induced 100% formation of friable callus cultivated under red-blue LED, demonstrating that the light quality significantly influenced callogenesis. Obtained results confirmed that in vitro regeneration and callogenesis is a useful strategy in the protection of endangered species. In this way, a new renewable source of biomass with high quality plant material is presented aiming at the bioprospecting of seedling extracts and friable callus to obtain secondary metabolites of this medicinal plant.

Download Full-text

Comparative Studies of In Vitro Regeneration Capacity in Some Breeding Forms of Prunus persica (L.) Batsch

BIO Web of Conferences ◽

10.1051/bioconf/20202400055 ◽

2020 ◽

Vol 24 ◽

pp. 00055

Author(s):

Irina Mitrofanova ◽

Nina Lesnikova-Sedoshenko ◽

Olga Mitrofanova ◽

Anatoliy Smykov ◽

Svetlana Chelombit

Keyword(s):

Prunus Persica ◽

Culture Media ◽

In Vitro Regeneration ◽

Regeneration Capacity ◽

Hybrid Form ◽

Murashige Skoog ◽

Morphogenic Callus ◽

High Regeneration ◽

Hybrid Forms

Peach [Prunus persica (L.) Batsch] is one of the most important stone fruit crops in the world. Preservation of valuable genotypes and creation of new breeding forms need the effective methods for plant propagation. Biotechnological method makes it possible to multiply valuable genotypes in vitro and produce high-quality plant material. Plantlets were obtained from hybrid peach embryos in five cross combinations. The induction of morphogenesis and the studies of regenerative capacity were carried out on culture media Murashige, Skoog (MS) and Gamborg, Eveleigh (B5) with vitamins and plant growth regulators. The segments of plantlets with 2-3 internodes were placed on MS and B5 media. Use of B5 medium with 0.75-1.0 mg L-1 BAP and 0.1 mg L-1 IBA induced organogenesis in the studied hybrid forms. The microshoots of the hybrid form ‘Summerglo’ × ‘Nikitskiy Podarok’ had a high regeneration capacity. In the forms ‘Persey’ × ‘Nikitskiy Podarok’ and ‘KAT 92-2210’ × ‘Nikitskiy Podarok’ low regeneration capacity was noted. An increase in BAP concentration resulted in formation of hydrated microshoots and non-morphogenic callus. It was determined that to obtain normal peach microshoots, the optimal culture parameters were a temperature of 24 ± 1oC, 16-hour photoperiod, and 37.5 μM m-2s-1 light intensity.

Download Full-text

Efficient plant regeneration via meristematic nodule culture in Paeonia ostii ‘Feng Dan’

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-021-02216-x ◽

2022 ◽

Author(s):

Li Xu ◽

Fangyun Cheng ◽

Yuan Zhong

Keyword(s):

Industrial Development ◽

Woody Plant ◽

In Vitro Regeneration ◽

Shoot Elongation ◽

Attractive Alternative ◽

Tree Peony ◽

Induction Rate ◽

Meristematic Nodules ◽

Propagation Methods

AbstractTree peony (Paeonia sect. Moutan) is an economically important multipurpose woody plant in terms of its medical, ornamental and oil values, but its breeding and industrial development are severely limited due to inefficient traditional propagation methods and existing in vitro regeneration systems. Meristematic nodules (MNs) are an attractive alternative to solve this problem. This study first presented a protocol for in vitro regeneration of P. ostii ‘Feng Dan’ via MN culture with four consecutive steps, including embryogenic callus (EC) formation, MN induction and leaf cluster differentiation, shoot elongation, rooting and acclimatization. The highest EC induction rate (81.25%) was achieved when cotyledons were cultured on modified Murashige and Skoog (mMS) medium with 4.04 µM N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU) + 5.37 µM α-naphthylacetic acid (NAA) for 30 days. The optimal MN induction rate (100%) and leaf cluster differentiation rate (45.83%) were obtained when ECs were cultured on modified woody plant medium (mWPM) supplemented with 2.02 µM CPPU + 2.27 µM thidiazuron (TDZ) for a subculture time of 10 days. The combination of 1.29 µM 6-benzyladenine (BA) + 0.58 µM gibberellin (GA3) yielded the best shoot elongation (13.40 shoots per nodule), rooting rate (43.33%) and consequently survival rate (45.83%). The study will be beneficial to the mass propagation, breeding and genetic improvement of tree peony.

Download Full-text