scholarly journals Badanie wzrostu kultur korzeniowych oraz tworzenia i wzrostu in vitro kalusów z różnych organów marchmi odmiany Terfekcja [Growth and callus formation of tissue cultures derived from various carrot organs]

2015 ◽  
Vol 29 (1) ◽  
pp. 25-42
Author(s):  
Anna M. Domańska ◽  
Aldona Rennert

The clones of excised roots, leaves, petioles, cotylenods, hypocotyls and root calluses derived from the respective carrot fragments (cv. 'Perfekcja' commonly cultivated in Poland) were cultured <i>in vitro</i>. An influence of thiamine concentrations on the growth of root tissue was examined. Several various media were tested for callus cultures. Bee bread extract was also applied. The growth of isolated clones during early and later culture periods was compared.

Author(s):  
K. Murugan ◽  
Ashok Kumar Sahoo ◽  
Bosco Lawarence,

To evaluate the anti-metastatic potentialities of purified anthocyanin from Osbeckia aspera (L.) Blume. and O. reticulata Bedd. against selected human cancer cell lines such as HT29 colon, MG63 bone and HeLa cervical by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis and DNA fragmentation test. Anthocyanin was extracted from the in vitro callus culture of the Osbeckia species, purified using amberlite column chromatography and fractionated by LC-MS/MS. Anthocyanin producing callus cultures were trialed on MS medium fortified with various combinations of phytohormones and sucrose. Significant callus formation in O. aspera was initiated in cultures containing 0.5 mg/L of 2, 4-D and 0.5 mg/L BA, while that in O. reticulata was initiated with 1.2 mg/L BA and 1.4 mg/L NAA. The same hormonal combinations on sub-culturing turned white friable callus into red compact callus. Purified anthocyanins obtained from O. aspera and O. reticulata contained Malvidin-3 -diglucoside, delphinidin, cyanindin aglycone and Peonidin. Osbeckia species displayed differential responses against the HT29 colon, MG63 bone and HeLa cervical cancer cell lines in terms of IC50 values of toxicity. O. aspera was more effective against HeLa cervical cell lines (23.7μg/ml) followed by HT29 colon (64.7μg/ml) as compared to O. reticulata. Poor selectivity index was noticed with bone cancer cell lines. The results were substantiated by apoptotic analysis and DNA fragmentation results. The overall results suggest that the purified anthocyanin of O. aspera and O. reticulata was excellent as antimetastatic and warrant further studies to isolate novel compounds for chemotherapeutic use.


2021 ◽  
Vol 1 (25) ◽  
pp. 98-112
Author(s):  
N.A. Yegorova ◽  
◽  
I.V. Stavtzeva ◽  

To increase the efficiency in agricultural plant breeding, including clary sage – one of the main essential oil crops grown in Russia, it is necessary to use biotechnological methods. One of these techniques is based on the induction of somaclonal variability in the callus tissue culture. To develop it, it is necessary to optimize the conditions for obtaining plant-regenerants in vitro and their analysis. The aim of this work was to study the features of morphogenesis and regeneration of plants from callus cultures to develop cell technologies for creating an initial breeding material based on somaclonal variability in Salvia sclarea L. In the course of the research, we found that the optimal explants for obtaining morphogenic callus, from which shoots were regenerated, were segments of buds and stems with a node (isolated from seedlings in vitro). Cytological analysis of callus cultures revealed two types of morphogenesis – organogenesis (gemmogenesis) and somatic embryogenesis. The features of the morphogenic callus formation of six sage cultivars and samples during the long-term cultivation were studied. The maximum frequency of morphogenesis was noted in the 2nd passage (from 32.4 to 85.2 %, depending on the genotype). Then, to the 8–10th passage, this indicator decreased to 0.0–3.9 %.‘S-785’ and ‘Taigan’ cultivars showed the highest morphogenesis frequency (81.5–85.2 %) and duration of callus regeneration potential (up to the 10th passage). The analysis of callus cultures of six donor plants of ‘S-785’ cultivar helped us to reveal their heterogeneity in morphogenesis induction ability. The maximum frequency of morphogenic callus formation (76.3–91.5 % in the 2nd passage) and the duration of the morphogenic potential preservation (up to the 12th passage) were observed in plants No. 3 and 9, whereas in No. 2, regeneration with a frequency of 3.6–9.7 % was observed only during three passages. Analysis of plants obtained from calli showed their variability in morphology – up to 12.5 % of the samples had deviations compared to the initial cultivar ‘S-785’ in leaf shape, inflorescence structure, flower color, etc. Somaclonal changes in morphological and economically useful traits revealed in regenerants indicate that they are promising for use in sage breeding.


2017 ◽  
Vol 59 (2) ◽  
pp. 31-39 ◽  
Author(s):  
Csongor Freytag ◽  
Sándor Attila Pabar ◽  
Zita Demeter ◽  
Ádám Simon ◽  
Anna Resetár ◽  
...  

AbstractWe aimed to produce tissue cultures and plant regeneration from endangered Crocus species: C. scepusiensis, C. tommasinianus, C. vittatus (“Verni” series of the genus) and C. banaticus. For initiation of cultures we used a plant growth regulator (PGR) combination used for in vitro culture of saffron and its relatives: 10 mg L-1 α-naphthaleneacetic acid (NAA) and 1 mg L-1 6-benzyladenine (BA). Shoot tips of young seedlings (C. scepusiensis) and corms (for the rest of species) were used as explants. C. scepusiensis explants developed into organogenic calli. On media with decreased NAA and with or without increased BA concentration, calli produced stigma-like structures and/or shoots and whole plants. In the other species, callus initiation medium induced callus formation with abundant somatic embryos. In C. tommasinianus, embryos developed shoots when auxin content of medium was decreased. In C. banaticus, a decrease of auxin with or without an increase in cytokinin content led to shoot or whole plant regeneration, as in C. scepusiensis. In the case of C. vittatus and C. banaticus, initiation and/or maintenance of cultures on indole-3-butyric acid (IBA) and increased sucrose concentration stimulated whole plant regeneration and in vitro cormlet development. C. scepusiensis and the rest of cultures (organogenic vs. embryogenic) differed at the biochemical level: C. scepusiensis cultures had higher (yet still low) enzymatic antioxidant (catalase, peroxidase) activities. With respect to catalase isoenzyme patterns, C. banaticus was different from the rest of cultures, demonstrating its distinct taxonomical position. Besides germplasm preservation use of the present cultures, they have a potential biotechnological value.


2021 ◽  
Vol 875 (1) ◽  
pp. 012082
Author(s):  
O S Mashkina ◽  
T M Tabatskaya ◽  
O M Korchagin

Abstract In vitro modelling of stress is one of the promising avenues for plant breeding for tolerance to negative environmental factors. In this study we examined the effect of NaCl (0.5%) on callusogenesis and morphogenesis of stem explants of different birch genotypes: Betula pendula Roth, B. pendula Roth var. carelica (Mercklin) Hämet-Ahti, B. pendula f. ‘dalecarlica’ (L.f.) Schneid., B. pubescens Ehrh. In our experiments we used pre-selected microclones from our in vitro collection on NaCl (0.2-1.0%) selective media. The clones were contrasted by the degree of their sensitivity to salinity (so-called ‘stable’ and ‘sensitive’ microclones). With the use of stem callus cultures we identified informative, simple and reproducible indicators for the selection of salt-tolerant genotypes. Among these indicators were the frequency of callus formation and the viability of callus cultures, which were significantly higher in ‘stable’ group of microclones. Polyploid birch clones (2n=4x=56, 2n=3x=42) were more resistant to salination compared to diploid clones (2n=28). Our study has shown that the selection of salt-tolerant birch lines can be based on the plants’ genetic diversity presented in the collection (various species, varieties, hybrids, polyploids) and manifested in the process of in vitro cultivation, as well as in the cellular heterogeneity of callus cultures.


Author(s):  
Noor Aini Habibah ◽  
Sukarti Moeljopawiro ◽  
Kumala Dewi ◽  
Ari Indrianto

<p><em>Stelechocarpus burahol </em>is one of the medicinal plants that contains flavonoids. The study was carried out to know flavonoid production of cultures in vitro <em>S. burahol </em>from mesocarp explants. Mesocarp explants were cultured on MS medium containing different combination and concentration of plant growth regulators i.e. picloram (5, 7.5 and 10 mg/L) and 2, 4-D (10, 15 and 20 mg/L) under dark condition. Induction of callus formation started on the 20.29<sup>th</sup> to the 29.86<sup>th</sup> days. Medium supplemented with Picloram and dark state proved to be the best condition for optimum callus induction from mesocarp explants of <em>S. burahol</em>. Callus grown on medium with the addition of 7.5 mg/l Picloram produces the highest flavonoid. The maximum production of the secondary metabolite was obtained from 8 weeks old callus. However, by the time of callus ageing, its output has declined. It could be concluded that callus cultures from mesocarp <em>S. burahol</em> can be used for flavonoid production. </p><p><strong>How to Cite</strong></p><p>Habibah, N. A., Moeljopawiro, S. Dewi, K. &amp; Indrianto, A. (2016). Flavonoid Production In Callus Cultures From Mesocarp <em>Stelechocarpus burahol</em>. <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(2), 214-221.</p>


ISRN Botany ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Alla I. Yemets ◽  
Galina Ya. Bayer ◽  
Yaroslav B. Blume

Efficient protocols for callus production, plantlet regeneration, protoplast isolation, and micronucleation of finger millet (Eleusine coracana (L.) Gaertn.) were developed. White nodulated calli were formed on medium with N6 macrosalts, MS microsalts, 2.4-dichlorophenoxyacetic acid (2 mg L−1), kinetin (0.4 mg L−1), 1-naphthalene acetic acid (2 mg L−1), and certain additives. It was found that appropriate supplementation leads to formation of numerous shoots. Healthy rooted plantlets formed on hormone-free media. Although different tested additives had no significant effect on percentage of callus formation, it affected callus quality that further dictated plant-forming capacities. Seedlings were better source tissues for protoplasts isolation compared to callus cultures. About protoplasts were isolated from one gram of seedling coleoptyles. Microcolonies were visible after 20–25 days' incubation on KM8p medium supplemented with glutamine (100 mg L−1) and proline (500 mg L−1). Here we also present a procedure of an efficient induction of micronuclei after chlorpropham (10 μM) and cytochalasin-B (20 μM) seedlings treatment with subsequent microprotoplasts isolation. This technique is discussed for the transfer of alien chromosomes and genes from finger millet by microprotoplast-mediated chromosome transfer.


2009 ◽  
pp. 1-15
Author(s):  
Maylin Pérez-Bernal ◽  
Magalis Delgado Rigo ◽  
Carlos Alberto Hernández Díaz ◽  
María Teresa Barceló Ávila ◽  
Raúl Armas Ramos

Most of Cuban rice cultivars are classified into indica subspecies, and they are inclined to poor in vitro response. In this paper we studied the role of endosperm and amino acids on callus formation of two Cuban rice cultivars: J-104 and IACuba-28. Callus cultures were initiated from three treatments for mature seed: intact seed, embryo with scutellum but without endosperm, and endosperm alone. It demonstrated the direct incidence of endosperm on in vitro seed contamination. But the higher percentage of embryogenic calli was obtained from intact seeds, despite of 12.94 % of seed contamination. Callus formation from endosperm alone did not occur. The role of endosperm to successful callus formation from scutellum was discussed. Effect of amino acids on rice callus growth from intact seeds was examined by supplying callus formation medium with glutamine and proline, separately or in combination, in both cultivars. Callus formation of J-104 was improved considerably with 500 mg/l of proline and glutamine in the culture medium, but in IACuba-28 were not observed significant changes. The percentage of embryogenic callus and the increase of fresh weight of calli were correlated with genotype and amino acid supplement in culture medium.


2018 ◽  
Vol 22 ◽  
pp. 274-281
Author(s):  
N. B. Kravets ◽  
N. V. Tulaidan ◽  
M. Z. Mosula ◽  
N. M. Drobyk

Aim. The aim of the research was to choose the conditions for microclonal propagation and obtain callus cultures from Carlina аcaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl plants in vitro. Methods. For microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia we used rosettes of 2–3-month specimens and planted them on semi-solid Murashige and Skoog (MS) medium with decreased macro- and microsalts concentrations (MS/2) supplemented with kinetin (Кin) (from 1–3 mg/l) and 0.1 mg/l of 1-naphthaleneacetic acid (NAA). For induction of callus formation, we used root, stem explants from С. acaulis, C. cirsioides and C. onopordіfolia, and planted them on nutrient media MS, MS/2, and Gamborg and Eveleigh (В5) supplemented with different concentrations of cytokinins – 6-benzylaminopurine (BAP) or Кin and auxins – 2.4-dichlorophenoxyacetic acid (2.4-D) or NAA and indole-3-acetic acid (IAA). Results. MS/2 medium supplemented with growth regulators of NAA and Кin were the most efficient to provide the formation of microclones. For C. сirsioides plants, this indicator was 6.6–6.8 rosettes per graft after 6 months of cultivation and for С. acaulis and C. onopordіfolia – 4.2–5.0 and 4.8–5.2 respectively. To raise the percentage of rooting for microclones of Carlina species, it was expedient to steep them preliminarily in the solution of indole-3-butyric acid (IBA) with 1000 mg/l concentration for a minute. Optimal for obtaining callus tissue from Carlina plants was nutrient medium MS supplemented with 3 mg/l IAA, 0.5 mg/l NAA and 0.5 mg/l Kin and MS/2 with 0.1 mg/l BAP and 0.5 mg/l 2.4-D; under such conditions the percentage of callus induction exceeded 90 % for all types of explants. Conclusions. There were chosen the conditions for microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia and worked out the schemes for enrooting obtained microclones in vitro. Capable of growing rapidly callus cultures from root and stem explants of the investigated plant species were obtained. Keywords: Carlina аcaulis L., Carlina cirsioides  Klok, Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl, in vitro, microclonal propagation, callus induction.


Genome ◽  
1987 ◽  
Vol 29 (1) ◽  
pp. 122-128 ◽  
Author(s):  
Michael Lee ◽  
R. L. Phillips

Chromosomal instability is a common occurrence in plant tissue cultures and has been documented in plants regenerated from several genotypes of maize (Zea mays L.) tissue cultures. The objective of this research was to evaluate the frequency and types of chromosomal aberrations in regenerated plants of an Oh43–A188 genetic background, which had not been examined previously for chromosome stability in culture. Organogenic callus cultures were intitated from immature embryos of F2 plants for several Oh43 ms isoline × A188 crosses. The chromosome constitution of 267 plants was investigated through meiotic analysis of plants regenerated either 3 to 4 or 8 to 9 months after culture initiation. No abnormalities were detected in 78 plants regenerated during the first period. During the second period, however, 91 of the 189 plants were cytologically abnormal. One hundred and eight aberrations were detected and most (96%) involved changes in chromosome structure such as interchanges (42%), deficiencies (35%), and heteromorphic pairs (19%). All deficiencies were intercalary. Also, most (51%) interchanges involved chromosome 6. An association between male-sterility factors and chromosome instability was not observed. Breakpoints were primarily on chromosome arms containing large blocks of heterochromatin such as knobs. Several abnormal plants from the same culture appeared to contain identical aberrations indicating the aberrations may trace to a single event. A hypothesis for the involvement of heterochromatin in chromosome breakage during in vitro culture is supported. Key words: Zea mays L., tissue culture, somaclonal variation, chromosome breakage, heterochromatin.


1976 ◽  
Vol 54 (7) ◽  
pp. 616-618 ◽  
Author(s):  
Charles W. Heuser ◽  
Darrel A. Apps

Plantlet regeneration has been induced from callus tissue cultures obtained from petal parts of Hemerocallis cv. Chipper Cherry. Callus cultures capable of regenerating whole plantlets were established on the agar-solidified Murashige and Skoog's medium supplemented with (2,4-dichlorophenoxy)acetic acid (2,4-D) (1.0 mg/litre) + kinetin (1.0 mg/litre). The callus formed was dense, yellowish-green in color, and had what appeared to be meristematic protuberances. Shoots and roots developed when the callus was subcultured on a medium lacking 2,4-D.


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