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1

Embryogenic callus formation by cotyledon and leaf explants of Crambe abyssinica seedlings

100%
Furmanek T., Banas W.
Biotechnologia
|
2011
|
issue 2
209-213
EN
The study investigates the regeneration capacity of Crambe abyssinica via the callus phase. The conducted experiments provide an overview of various conditions appropriate for callus development and its subsequent differentiation. Explants from cotyledons and leaves of sterile 14-day-old seedlings of C. abyssinica cv. Mayer were used. The primary callus so obtained was subsequently utilised for the observation of embryogenesis. The best callus formation on the explants was achieved on the agar-solidified MS medium with 0.5 mg . dm-3 NAA and 0.5 mg . dm-3 BAP or TDZ. The formation of secondary embryogenic callus was most efficient on MS medium supplemented with 0.4 mg . dm-3 NAA and 0.3 mg . dm-3 of BAP.
2

Suspension cultures of Pharbitis nil

100%
Szyp I., Tretyn A.
Biotechnologia
|
2001
|
issue 1
127-130
EN
Suspension cultures are more suitable for physiological, biochemical and molecular investigations than callus cultures grown on solid media, because the former provide more homogeneous system than the latter ones. A large number of plants were found suitable for establishing cell suspension cultures. In this study we describe the obtaining of cell suspension cultures derived from callus induced from cotyledons of Pharbitis nil. Explants isolated from plants grown under inductive and non-inductive conditions were cultured on MS basal medium containing various concentrations and combinations of growth regulators. To initiate the cell suspension culture, small clumps of friable callus obtained from cotyledons were suspended in liquid callusing medium. An initial inoculum density was 2104cells/ml. Every 20 days the cultures were transferred to a fresh medium. The cell number in suspension was determined by direct microscopic counting with haemocytometr. The cell suspension cultures contained both single cells and small cell aggregates.
3

Estimation of callus formation capability of anthers obtained as a result of crossing Linola cultivar with other linseed cultivar plants

100%
Wielgus K., Mankowska G., Bialas W.
Biotechnologia
|
2008
|
issue 2
84-89
EN
Identification of responsive genotypes and development of an efficient and cost-effective doubled haploid production system in linseed (Linum usitatissimum L.) is required for application in practical breeding program. Although the technique has been improved in recent years, there is limited data on the inheritance of callogenesis and plant regeneration in linseed anther culture. The objective of this research was to estimate the influence of the ?Linola' cultivar genome and other cultivars used for crossing with 'Linola' plants in a process declining during the anther cultures in F1 generation. 3144 anthers altogether were placed on the medium, on average 393 from each population. Hybrid lines obtained as a result of crossing 'Linola' x 'A.C. Emmerson' were not capable to form callus. The anthers from crossing of 'Linola' x 'Szafir', 'Linola' x 'Opal' and 'Linola' x 'Bionda' were characterized with higher than average capacity to form callus, while the anthers from crossing of 'Linola' x 'Norlin', 'Linola' x 'Vinny' and 'Linola' x 'Oliwin' had lower potential of forming callus.
4

The androgenic response of Capsicum frutescens L. genotypes in in vitro anther culture

100%
Nowaczyk P., Kucharska A., Nowaczyk L.
Biotechnologia
|
2007
|
issue 1
197-202
EN
Two soft-flesh Capsicum frutescens L. lines were used in the study. The androgenesis process was induced according to the procedure described for C. annuum L. In the cultures established in summer, not many anthers forming the callus tissue were observed. Cytometric analysis showed the presence of cells with different content of DNA. Among 1.000 anthers displayed of one line, seven embryos were observed. Four embryos continued the development resulting in obtaining plants. Three of them had the DNA content on 1C level, the fourth one was diploid. The test of an androhaploid induction in autumn cycle, with the genotype of positive response in summer, was not successful.
5

The effect of cytokinins on morphogenetic response of Polemonium coeruleum seedlings explants

100%
Trejgell A., Tretyn A.
Biotechnologia
|
2010
|
issue 2
125-131
EN
The aim of the presented research was to examine the morphogenetic response of Polemonium coeruleum explants. The donor material were 10-day-old seedlings. Surface sterilized seeds were germinated on MS medium supplemented with GA3 (1 mg?dm-3). Seedling explants (shoot tips, fragments of cotyledons, hipocotyls and roots) were isolated and transferred onto solidified MS medium supplemented with different types of cytokinins (BA, KN, ZEA, 2iP) at concentrations 1.0, 3.0 and 5.0 mg?dm-3 in combination with NAA (0.1 mg?dm-3). All explant types were characterized by callus proliferation. It was observed that calli developed on the entire surface of hipocotyl and root fragments. On the other hand, shoot tips and cotyledonary petioles formed callus tissue at the cut ends, and petioles only at abaxial ends. The growth of calli on all explant types was strongly stimulated by ZEA. Among the explants tested, only shoot tips exhibited shoot organogenesis. The highest frequency of shoot organogenesis was observed when the explants were cultured on a medium supplemented with 5.0 mg?dm-3 BA (100%) or 5.0 mg?dm-3 ZEA (97%). The highest shoot number per explant was obtained in the presence of 5.0 mg?dm-3 ZEA (8.4 on average). The presence of BA or ZEA in the proliferation medium inhibited rhizogenesis and the elongation growth of shoots. However, root organogenesis was supported by KN added into the medium.
6

Anthocyanis in suspension cultures orginating from different parts of sweetpotato plant

100%
Konczak-Islam I., Yoshinaga M., Nakatani M., Yamakawa O.
Biotechnologia
|
1999
|
issue 3
143-153
EN
Anthocyanin accumulating cell lines were established from storage root, leaf and stem explants of sweetpotato (Ipomoea batatas (L.) Lam.), cv. Ayamurasaki. The calli developed on MS basal medium enriched with 2,4-D at 27?C accumulated pigments in the dark. The storage root originated suspension culture generated the highest amount of total anthocyanins expressed as colour value (5.9) followed by the cultures originating from leaf (4.3) and stem (1.7). The cultures displayed similar anthocyanin profile regardless of source of explants. The major pigments derived from suspension cultures appeared with earlier retention time on ODS-column HPLC than the YGM pigments accumulated in vivo, which suggests that they are highly hydrophilic and have simpler chemical structure.
7

Effect of selected factors on the effectiveness of Capsicum annuum L. anther culture

88%
Nowaczyk P., Kisiala A.
|
EN
The primary aim of the study was to establish the effectiveness of induced androgenesis in in vitro anther culture of two pepper (Capsicum annuum L.) breeding lines ? ATZ1 and PO, and a hybrid between these two lines (ATZ1 ? PO)F1. Anther culture was maintained according to the method developed by Dumas de Vaulx et al. (1981) with some modifications. The experiment revealed that the effectiveness of androgenesis ranged from 4% for the ATZ1 line to 1.5% for the (ATZ1 ? PO)F1 and strongly depended on the developmental stage of flower buds, as well as the conditions for anther culture maintenance. The development of androgenic embryos was successfully induced only in anthers which originated from the flower buds with petals equal or slightly longer than sepals and there was a clear relationship between the length of the period of anther induction on CP medium and the level of kinetin in R1 regeneration medium.
8

The effects of 2,4-D, NAA and picloram on callus induction and somatic embryogenesis in ornamental asparagus

88%
Kepczynski J., Zielinska S., Kepczynska E.
Biotechnologia
|
2001
|
issue 2
170-173
EN
In this study we examined the effect of 2,4-D, NAA and picloram at 1.5, 3.0 and 4.5 mg/l in MS medium on callus induction and somatic embryogenesis in Asparagus densiflorus cv. Sprengeri and Asparagus plumosus. The callus formation of both shoot tips and nodal explants from A. densiflorus after 8 weeks of culture on MS medium supplemented with NAA and picloram (1.5, 3.0 mg/l) was observed. However, callus on medium containing 2,4-D was occasionally induced. On medium supplemented with picloram shoot tip explants of Asparagus plumosus showed callus inducing capacity. After four weeks of callus culture on hormone-free medium the globular embryos were achieved. On callus induced NAA organogenesis was observed.
9

The effect of cefotaxime and carbenicillin on viability and development of carrot protoplasts

88%
Grzebelus E., Grabka M., Baranski R.
Biotechnologia
|
2010
|
issue 2
156-164
EN
We examined the toxicity of two antibiotics belonging to the betalactam group, to carrot (Daucus carota L.) protoplasts. Leaf protoplasts were cultured in the presence of cefotaxime or carbenicillin applied in five concentrations in the range from 0.1 to 0.5 mg ml-1. Cell viability, division frequency, and regeneration capacity were assessed to determine the potential toxic effect of the antibiotics. Both antibiotics significantly reduced protoplast viability and their ability to divisions. Their toxic effect intensified linearly with increasing antibiotic concentrations in the culture medium. More pronounced negative effect exhibited carbenicillin, which was evident 24 h after protoplast isolation. It also lowered cell mitotic activity two- to ten-fold, as compared to the control. Despite different reaction of cells exposed to carbenicillin and cefotaxime, callus tissue and somatic embryos were successfully obtained and allowed efficient plant regeneration. The comparison of the obtained results indicates that cefotaxime used in concentrations up to 0.2 mg ml-1 can be recommended in carrot cell cultures to prevent microbial contamination.
10

The effect of cytokinins on in vitro morphogenesis of sugar beet (Beta vulgaris L.)

88%
Goska M.
Biotechnologia
|
2004
|
issue 2
137-145
EN
The aim of the studies was to estimate the effect of cytokinins on the production of callus and shoots from different seedling explants of sugar beet. Two diploid multigerm genotypes were used in the experiment. The seedling explants (hypocotyls, cotyledons and leaves) were cultured on MS medium supplemented with cytokinins: BAP, 2iP, TDZ in concentrations 0,2; 1,0 and 2,0 mg/l. Hypocotyls, cotyledons and leaves showed different abilites to form callus and shoots. After two weeks of culture, the induction of callus was mainly observed on the explants of hypocotyls and cotyledons, whereas adventitious shoots formed on leaf explants. Frequency of callus and shoots induction was dependent on the origin of the explants, concentration and type of the cytokinins. The highest percentage of regeneration was obtained on the explants of young leaves, on medium containing TDZ (51,5%). The optimal concentration of cytokinins was 1 mg/l. Plant regeneration on seedling explants came only under direct organogenesis.
11

Cell suspension obtained from callus or directly from cotyledons of Pharbitis nil Chois

88%
Trejgell A., Wisniewska J., Tretyn A.
Biotechnologia
|
2004
|
issue 2
219-224
EN
An attempt has been made to obtain cell suspension from callus as well as directly from cotyledons P. nil. Cotyledons of 7-days old sterile seedlings and flower buds excised from 3-week old plants were the material for the induction of callus. The explants were laid out on MS medium supplemented with various combination of plant hormones: BAP (5 mg/l) and NAA (1 mg/l) and supplemented with 3% sucrose or 2% glucose and 1% sucrose, BAP (5 mg/l) and IAA (0,5 mg/l), BAP (0,5 mg/l) and Picloram (1 mg/l), BAP (5 mg/l) and Picloram (0,5 mg/l), 2,4-D (0,125 mg/l).The cultures were grown in continuous white light or in darkness. The callus obtained from cotyledons cultivated in darkness and callus from flower buds cultivated in light on MS medium with BAP (5 mg/l), NAA (1 mg/l), 2% glucose and 1% sucrose were proved useful for obtaining cell suspension. Moreover, an attempt was made to obtain cell suspension directly from cotyledons. Cotyledons were cut into small fragments or were subjected to enzymatic digestion. The cell suspensions were cultivated on MS medium with the addition of BAP (5 mg/l) and IAA (0,5 mg/l) on a shaker at 140 rpm. The increase of cell number was determined by counting the cells every 5 days. In the subsequent subcultures, a decrease of the number of cell divisions was observed.
12

Instability of ploidy level during regeneration of transformed and non-transformed tobacco (Nicotiana tabacum L.)

75%
Sliwinska E., Zimny J., Drozdowska J.
|
2003
|
issue 3
260-266
EN
In plant tissue cultures, somaclonal variation is often observed. It can be an effect of the changes in the individual chromosome number or in the ploidy level. Flow cytometry, a fast and accurate method for the estimation of the nuclear DNA content, can be applied to study these changes. The DNA content in differentiated tissues of Nicotiana tabacum cultured in vitro was estimated using Partec CCA flow cytometer, starting from explant, through callus, up to regenerated shoots. The explant constituted stem segments of N. tabacum plants, non-transformed and transformed with gfp gene. Flow cytometric analysis showed differences in the proportion of 2C, 4C, 8C and 16C cells in plant tissue in different culture stages. Among the regenerated plantlets originated from non-transformed and transformed plants, diploid, tetraploid and mixoploid forms were observed. The transformation did not influence the share of cells representing different ploidy levels in the investigated plant material.
13

Densytometric analysis of diosgenin in extracts from callus tissue of Polygonatum verticillatum (L.) All.

75%
Szybka-Hrynkiewicz P., Janeczko Z.
Biotechnologia
|
2004
|
issue 2
93-99
EN
In this study, we examined the effect of plant growth regulators (2, 4-D, 6-BAP) and steroidal hormone (17-beta-estradiol) on a quantity of diosgenin in callus tissue of Polygonatum verticillatum (L.) All. The callus grown in in vitro conditions on MS medium with: 1/1, 2/2, 5/5, 10/10 ([mg/dm3] 2,4-D/[mg/dm3] 6-BAP) and 1/1/1, 2/2/2, 5/5/5, 10/10/10 ([mg/dm3] 2,4-D/[M/dm3] 17beta-estradiol/[mg/dm3] 6-BAP) was used for the densytometric analysis to determine the presence of diosgenin. The content of diosgenin ranged from 0,0642 to 1,1458%. An increase in diosgenin content due to addition of 6-BAP and 2,4-D was observed. The developed procedure was estimated by statistical analysis which indicates that it can be used for routine analysis.
14

Agrobacterium-mediated transformation of yellow lupin to generate callus tissue producing HBV surface antigen in a long-term culture

75%
Pniewski T., Kapusta J., Plucienniczak A.
Journal of Applied Genetics
|
2006
|
vol. 47
|
issue 4
309-318
EN
The idea of an oral vaccine administered as a portion of plant tissue requires a high level of antigen production. An improved protocol for the induction of transgenic yellow lupin calli or tumours, reaching 44% of transformation rate, is presented here. It has been developed by using the nptII marker gene and the uidA reporter gene as well as various Agrobacterium strains and plant explants. This method of seedling and hypocotyl transformation was applied to raise calli or tumours producing a small surface antigen of Hepatitis B Virus (S-HBsAg). Lupin tissue lines were long-term cultured on selection media maintaining the growth rate and high expression level of the native form of S-HBs, up to 6 mug per g of fresh tissue.
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