OPTIMIZATION OF THE CONDITIONS OF ADAPTATION OF REGENERATED PLANTS OF BLACK CURRANTS (RIBES NIGRUM L.) VARIETIES BASHKIR BREEDING IN THE TRANSLATION FROM IN VITRO TO EX VITRO

2019 ◽  
Vol 0 (1) ◽  
pp. 83-88 ◽  
Author(s):  
L.A. Golovina ◽  
◽  
M.M. Ishmuratova ◽  
Keyword(s):  
Ribes Nigrum ◽  
Ex Vitro ◽  
Regenerated Plants

Long-term in vitro Storage Technique of Valuable Birch Genotypes and Plant Production on its Basis

2019 ◽  
pp. 57-67
Author(s):  
T.M. Tabatskaya ◽  
N.I. Vnukova
Keyword(s):  
Plant Production ◽  
High Survival ◽  
Ex Vitro ◽  
Regenerative Capacity ◽  
In Vitro Storage ◽  
Interspecific Differences ◽  
Regenerated Plants ◽  
Growing Conditions

A technique for the long-term (up to 27 years) in vitro storage of valuable birch genotypes under normal (25 °C, 2.0 klx, 16-h day and 8-h night) and low temperature (4 °C, 0.5 klx, 6-h day and 18-h night) growing conditions on hormone-free media has been described. The study explored for the first time the influence of different strategies to store the clones of Betula pubescens and B. pendula var. сarelica (6 genotypes) on the regenerative capacity of collection samples, adaptive potential of regenerated plants and plant production by the in vitro and ex vitro techniques. It was established that both storage strategies provided a persistently high survival rate (82-100%) and regenerative capacity of in vitro shoots (the multiplication coefficient of 4.2-6.3 and rhizogenic activity of 90-100%). The clones retained their characteristics of height growth under the in vitro and ex vitro conditions, and demonstrated intraclonal homogeneity and lack of signs of somaclonal variability. The plants showed substantial interspecific differences at the stage of multiplication and transfer to the greenhouse. The highest percentage of acclimated plants (75-98% depending on the clone genotype) was obtained after planting of micro plants straight in the greenhouse, which simplified the technology and made plant production less costly. long-term in vitro storage, birch, species, genotype, micropropagation, ex vitro adaptation, plant material

scholarly journals Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique

2013 ◽  
Vol 82 (3) ◽  
pp. 231-236 ◽  
Author(s):  
Maria Surma ◽  
Tadeusz Adamski ◽  
Wojciech Święcicki ◽  
Paweł Barzyk ◽  
Zygmunt Kaczmarek ◽  
...  
Keyword(s):  
Root Development ◽  
Ms Medium ◽  
Ex Vitro ◽  
Single Seed Descent ◽  
Yellow Lupin ◽  
Single Seed ◽  
Temperature Regimes ◽  
Regenerated Plants ◽  
Almost All

The aim of the studies was to establish in vitro conditions for the culture of pea and lupin embryos as the first step in the development of an in vitro assisted single seed descent technique for the attainment of homozygous populations. Materials for the study included of pea, and narrow-leafed and yellow lupin cultivars. Embryos dissected from mature but still-green seeds were cultured in vitro on two modified MS media and under three temperature regimes. Shoot and root lengths of regenerated plants were measured after 7, 14 and 21 days of culture. For pea plants full-strength MS medium with 4 g l<sup>−1</sup> agar and temperature 22/ 20°C (day/night) appeared to be the most conducive to shoot and root development, whereas for lupin plants lower temperatures were more propitious: 12°C in the dark for narrow-leafed lupin and 16/ 12°C (day/night) for yellow lupin. Almost all the cultured embryos developed into plants, but not all the regenerated plants survived acclimation to ex vitro conditions.

scholarly journals Droplet vitrification technique for cryopreservation of a large diversity of blackcurrant (Ribes nigrum L.) cultivars

2020 ◽  
Author(s):  
Saija Rantala ◽  
Janne Kaseva ◽  
Anna Nukari ◽  
Jaana Laamanen ◽  
Saara Tuohimetsä ◽  
...  
Keyword(s):  
Cultural Values ◽  
Core Collection ◽  
Gene Bank ◽  
Ribes Nigrum ◽  
Ex Vitro ◽  
Droplet Vitrification ◽  
Successful Transfer ◽  
Future Utilization

AbstractThe aim of plant gene banks is to preserve genetic resources selected based on their phenotypic, agronomic, historical or other cultural values for future utilization. In the present study the modified PVS2 droplet vitrification technique was tested and optimized for cryopreservation of a large diversity of blackcurrant (R. nigrum L.) accessions propagated in vitro and selected into a national gene bank core collection. Out of four accessions tested to optimize the method, three recovered and regenerated by 89–97% on average, but one recalcitrant in vitro line only by 25%. The tested post-cryopreservation recovery media with different macronutrient and growth regulator levels showed no generalized effect on regenerated shoots, but the effect of recovery media was different between cultivars. When the whole regeneration chain from cryopreservation via micropropagation to greenhouse conditions was tested, shoots at least 1 cm in length were found necessary for successful transfer ex vitro. The long-term cryopreservation of 22 blackcurrant accessions was finally conducted, with practices slightly modified from the tested protocol. The estimated recovery of shoot tips after 9 weeks in vitro was 17–94% with at least 75% recovery in seven accessions and at least 40% recovery in 19 out of 22 accessions. Only one accession had no cryopreservation success. The results demonstrated that the modified droplet vitrification technique is appropriate for a large diversity of blackcurrant accessions. However, cultivar-related differences and recovery procedures are to be considered for success in regeneration and ex vitro adaptation.

scholarly journals Assessment of Genetic Stability on In Vitro And Ex Vitro Plants of Ficus Carica Var. Black Jack Using Issr and Damd Markers

2021 ◽  
Author(s):  
Ankita Rajendra Parab ◽  
Chew Bee Lynn ◽  
Sreeramanan Subramaniam
Keyword(s):  
Stability Analysis ◽  
Genetic Stability ◽  
Similarity Index ◽  
Issr Markers ◽  
Ficus Carica ◽  
Ex Vitro ◽  
Regenerated Plants ◽  
Inter Simple Sequence Repeats ◽  
Dna Primers

Abstract In vitro propagation has been significant in producing a large number of genetically stable regenerated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium (WPM) supplemented with 20 µM 6-Benzylaminopurine (BAP) and 8 µM Indole-3-acetic acid (IAA) under different light treatments such as normal fluorescent white light (60 µmol.m− 2.s− 1), and four different LED spectra, white (400– 700nm), blue (440nm), red (660nm) and blue + red (440nm + 660nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack. Ten (10) primers of each ISSR and DAMD molecular marker were used to assess the genetic stability of the eight (8) samples of Ficus carica var. Black Jack, acquired over two years. The findings of this study revealed that inter simple sequence repeats (ISSR) and directed amplification of minisatellite DNA (DAMD) markers (DNA primers) are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR – DNA primer which was negated under the genetic similarity index analysis for the eight samples. It is recommended that genetic stability analysis should be performed for long-term maintenance of micropropagated plants.

scholarly journals Alkaloids of Seeds, in Vitro Cultivated, and ex vitro Adapted Plants of the Bulgarian Endemic Species Papaver Degenii (Papaveraceae)

2017 ◽  
Vol 12 (3) ◽  
pp. 1934578X1701200
Author(s):  
Tsvetelina Doncheva ◽  
Marina Stanilova ◽  
Vassil Vutov ◽  
Stefan Philipov
Keyword(s):  
Ex Vitro ◽  
Alkaloid Composition ◽  
Green Biotechnology ◽  
Aerial Parts ◽  
Alpine Zone ◽  
Regenerated Plants ◽  
In Vitro Shoots ◽  
Data Book ◽  
Glacial Relict

Papaver degenii (Urum. & Jav.) Kuzmanov is a Bulgarian endemic and glacial relict growing in the alpine zone of Pirin Mountains and it is included in the Red Data Book of Bulgaria. Numerous plants were in vitro multiplied in order to study the alkaloid composition of the species and to test the potential for green biotechnology establishment. Percentages of the crude alkaloid mixture in dried in vitro shoots and ex vitro adapted plants to room phytotron were similar to that in seeds (0.49%) and 5 to 6 times higher than in the aerial parts of wild-growing plants. Amurensine was the only alkaloid detected in the seeds. Four isoquinoline alkaloids were isolated from the regenerated plants, belonging to two alkaloid types: isopavines (amurensine and O-methylthalisopavine) and protopines (allocryptopine and protopine), amurensine being the main alkaloid: 63.4% in in vitro shoots, up to 88.1% in the aerial parts of ex vitro adapted plants. Allocryptopine was in higher amount (30%) in the roots of the ex vitro adapted plants.

scholarly journals Rapid In Vitro Propagation of Aruncus `Misty Lace', a New Heat-tolerant, Dwarf, Hybrid Goatsbeard

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 848A-848
Author(s):  
Hazel Y. Wetzstein* ◽  
Allan M. Armitage ◽  
Gwen N. Hirsch ◽  
Stephanie L. Anderson
Keyword(s):  
Plant Material ◽  
In Vitro Propagation ◽  
Shoot Organogenesis ◽  
Ornamental Plant ◽  
Naphthalene Acetic Acid ◽  
Ex Vitro ◽  
Regenerated Plants ◽  
Half Strength ◽  
Propagation Methods

Tissue culture is a useful means to clonally propagate new ornamental plant selections, particularly when plant material is limited and/or conventional propagation methods are ineffective. An efficient in vitro multiplication protocol was established to propagate a new goatsbeard hybrid (Aruncus dioicus, × A. aethusifolia). The hybrid is of interest because it exhibits a dwarf habit, delicate white flower panicles and fern-like leaves, yet is tolerant to heat and humidity. Experiments were conducted to evaluate explant type (nodes, stems, leaves, and floral parts), disinfestation procedures, and media formulations including varying concentrations of 6-benzylaminopurine (BAP) and naphthalene acetic acid (NAA). Rapid plant regeneration was obtained with a shoot organogenesis system using a half strength Murashige and Skoog medium supplemented with 4.4 μmol BAP, 0.54 μmol NAA, 30 g·L-1 sucrose, and 3.0 g·L-1 GelGro. Studies compared the performance and yield of plants rooted using different in vitro and ex vitro methods. Ex vitro rooting of shoots during greenhouse acclimatization under mist was most effective. Regenerated plants exhibited uniform and rapid growth, and performed well in greenhouse and field evaluations.

scholarly journals Organogenesis and long-term micropropagatlon of Polish pea cultivars

2011 ◽  
Vol 72 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Tomasz Pniewski ◽  
Joanna Wachowiak ◽  
Józef Kapusta ◽  
Andrzej B. Legocki
Keyword(s):  
De Novo ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Ex Vitro ◽  
Regenerated Plants ◽  
Axillary Meristems ◽  
Half Strength

The complete protocol for regeneration and long-term micropropagation of several Polish cultivars of pea (<em>Pisum sativum </em>L.) has been elaborated. The shoots were the most likely regenerated via de novo organogenesis. The adventitious buds formed in callus derived from cotyledons tissue adjacent to the axillary meristems of immature embryos. All cultivars' calli regenerated several shoots per explant on the MS medium supplemented with B5 vitamins and 4.5 mgl<sup>-1</sup> of BAP, however some differences in regeneration capacity among cultivars were observed. The plantlets were subsequently micropropagated with slightly higher efficiency and preserving a good viability over the long-term culture on a medium containing 2.0 mgl<sup>-1</sup> than one with 4.5 mgl<sup>-1</sup> of BAP. The additional step of the pre-conditioning culture of multiplicated shoots on a medium with very low BAP concentration i.e. 0.02 mgl<sup>-1</sup> was applied and appeared to be beneficial before rooting in vitro or grafting. The modified MS-derived medium with the half-strength of MS macroelements but with the full original dose of calcium and supplemented with B5 vitamins and 1.0 mgl<sup>-1</sup> of NAA was developed for effective rooting. The shoots were also sufficiently transferred into ex vitro conditions using grafting. The majority of the regenerated plants had adapted to in vivo conditions in a greenhouse and subsequently has set seeds. The presented protocol provides relatively efficient rate of de novo pea regeneration and would be useful for <em>Agrobacterium</em>-mediated transformation purposes.

Patrones de crecimiento de Cinchona officinalis in vitro y ex vitro; respuestas de plántulas micropropagadas y de semillas.

2017 ◽  
Vol 35 (1-2) ◽  
pp. 73-82
Author(s):  
Carlos Iván Espinosa ◽  
Gabriel Ríos
Keyword(s):  
Ex Vitro

El uso de herramientas biotecnológicas como la micropropagación se constituye en una alternativa de reproducción de especies amenazadas y con tamaños poblacionales reducidos. Sin embargo, uno de los problemas críticos en el uso de la micropropagación como herramienta de reproducción es la calidad de las plántulas resultantes en cuanto a su crecimiento y vigor. En el presente trabajo se evalua los efectos de la micropropagación sobre los patrones de crecimiento y sobrevivencia de plán­tulas in vitro de Cinchona officinalis L., una especie que ha sido fuertemente impactada por procesos de tala dentro de bosques naturales durante la época de la colonia. Se realizó un monitoreo de un total de 120 plántulas in vitro y 1988 plántulas ex vitro por 8 meses a partir del último repique. Adi­cionalmente, en cada plántula se contabilizó la cantidad de brotes axilares. Los resultados obtenidos mostraron un efecto remanente de los procesos de micropropagación, los cuales inicialmente inciden en la cantidad de brotes de las plántulas y en el crecimiento; sin embargo, este efecto no influye de forma negativa en la sobrevivencia de las plántulas durante la fase ex vitro

scholarly journals Metabolic profiling, in vitro propagation, and genetic assessment of the endangered rare plant Anarrhinum pubescens

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Asmaa Abdelsalam ◽  
Ehab Mahran ◽  
Kamal Chowdhury ◽  
Arezue Boroujerdi
Keyword(s):  
In Vitro Propagation ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Wild Plant ◽  
Rare Plant ◽  
Chemical Profiling ◽  
Nodal Cutting ◽  
Regenerated Plants ◽  
Plant Shoots

Abstract Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.

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